Tobacco inbred plants k326 src, cms k326 src, k346 src, cms k346 src, nc1562-1 src, nctg-61 src, cms nctg-61 src and hybrid nc196 src

ABSTRACT

The present disclosure provides tobacco inbred plants K326 SRC, CMS K326 SRC, K346 SRC, CMS K346 SRC, NC1562-1 SRC, NCTG-61 SRC, CMS NCTG-61 SRC and hybrid NC196 SRC. The present disclosure also provides parts of such plants and products made from those parts. The present disclosure also includes progeny of the provided plants including hybrids.

STATEMENT OF PRIORITY

This application claims the benefit, under 35 U.S.C. §119 (e), of U.S.Provisional Patent Application No. 61/751,720, filed on Jan. 11, 2013,and U.S. Provisional Patent Application No. 61/830,908, filed on Jun. 4,2013 in the United States Patent and Trademark Office the entirecontents of each of which is incorporated by reference herein.

INCORPORATION OF SEQUENCE LISTING

The Sequence Listing is hereby incorporated by reference in itsentirety, including the file named P34035US00.txt, which is 38,002 bytesin size and was created on Jan. 10, 2013, which is likewise hereinincorporated by reference in its entirety.

FIELD

The present disclosure provides tobacco inbred plants K326 SRC, CMS K326SRC, K346 SRC, CMS K346 SRC, NC1562-1 SRC, NCTG-61 SRC, CMS NCTG-61 SRCand hybrid cultivar NC196 SRC. The present disclosure also providesparts of such plants and products made from those parts. The presentdisclosure also includes progeny of the provided plants includinghybrids.

BACKGROUND

Tobacco (Nicotiana tabacum L.) is an important commercial crop in theUnited States as well as in other countries. In tobacco plants,N-demethylation of nicotine results in nornicotine, a secondary alkaloidknown to be a precursor for formation of N-Nitrosonornicotine (“NNN”) incured leaves. NNN is an undesired component of cured leaves.

The predominant alkaloid found in commercial tobacco varieties isnicotine, typically accounting for 90-95% of the total alkaloid pool.The remaining alkaloid fraction is comprised primarily of threeadditional pyridine alkaloids: nornicotine, anabasine, and anatabine.Nornicotine is generated directly from nicotine through the activity ofthe enzyme nicotine N-demethylase. Nornicotine usually represents lessthan 5% of the total pyridine alkaloid pool, but through a processtermed “conversion,” tobacco plants that initially produce very lowamounts of nornicotine give rise to progeny that metabolically “convert”a large percentage of leaf nicotine to nornicotine. In tobacco plantsthat have genetically converted (termed “converters”), the greatmajority of nornicotine production occurs during the senescence andcuring of the mature leaf (Wernsman and Matzinger (1968), Tob. Sci.,12:226-228). Burley tobaccos are particularly prone to geneticconversion, with rates as high as 20% per generation observed in somecultivars.

During the curing and processing of the tobacco leaf, a portion of thenornicotine is metabolized to the compound NNN, a tobacco-specificnitrosamine (TSNA) that has been asserted to be carcinogenic inlaboratory animals (Hecht and Hoffmann (1990), Cancer Surveys,8:273-294; Hoffmann et al, (1994), J. Toxicol. Environ. Health, 41:1-52;Hecht (1998), Chem. Res. Toxicol., 11:559-603). In flue-cured tobaccos,TSNAs are found to be predominantly formed through the reaction ofalkaloids with the minute amounts of nitrogen oxides present incombustion gases formed by the direct-fired heating systems found intraditional curing barns (Peele and Gentry (1999), “Formation ofTobacco-specific Nitrosamines in Flue-cured Tobacco,” CORESTA Meeting,Agro-Phyto Groups, Suzhou, China). Retrofitting these curing barns withheat-exchangers virtually eliminated the mixing of combustion gases withthe curing air and dramatically reduced the formation of TSNAs intobaccos cured in this manner (Boyette and Hamm (2001), Rec. Adv. Tob.Sci., 27:17-22.). In contrast, in the air-cured Burley tobaccos, TSNAformation proceeds primarily through reaction of tobacco alkaloids withnitrite, a process catalyzed by leaf-borne microbes (Bush et al. (2001),Rec. Adv. Tob. Sci., 27:23-46). Thus far, attempts to reduce TSNAsthrough modification of curing conditions while maintaining acceptablequality standards have not proven to be successful for the air-curedtobaccos.

SUMMARY

In an aspect, the present disclosure includes a seed of tobacco cultivarK326 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No. PTA-13455.

In another aspect, the present disclosure includes a tobacco plant, orpart thereof, produced by growing the seed of tobacco cultivar K326 SRC,a representative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13455.

In a further aspect, the present disclosure includes a harvested leaf,or part thereof, of a tobacco plant, or part thereof, produced bygrowing the seed of tobacco cultivar K326 SRC, a representative sampleseed of the cultivar having been deposited with the ATCC under ATCCAccession No. PTA-13455.

In an aspect, the present disclosure includes a harvested leaf, or partthereof, of a tobacco plant, produced by growing the seed of tobaccocultivar K326 SRC, a representative sample seed of the cultivar havingbeen deposited with the ATCC under ATCC Accession No. PTA-13455, wherethe leaf has a reduced amount of nornicotine and/orN′-nitrosonornicotine (NNN).

In an aspect, the present disclosure includes a harvested leaf, or partthereof, produced by growing the seed of tobacco cultivar K326 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13455, where the leaf has areduced amount of nornicotine and/or N′-nitrosonornicotine (NNN), wherethe reduced amount of nornicotine and/or N′-nitrosonomicotine (NNN) isreduced in a smoke stream produced from the leaf.

In a further aspect, the present disclosure includes a tobacco product,prepared from a tobacco plant, or part thereof, produced by growing theseed of tobacco cultivar K326 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13455, where the product is selected from the group consisting ofpipe tobacco, cigar tobacco, cigarette tobacco, chewing tobacco, leaftobacco, shredded tobacco, and cut tobacco.

In an aspect, the present disclosure includes a tobacco product,prepared from a tobacco plant, or part thereof, produced by growing theseed of tobacco cultivar K326 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13455, where the product is selected from the group consisting of acigarillo, a non-ventilated recess filter cigarette, a vented recessfilter cigarette, a cigar, snuff, and chewing tobacco.

In another aspect, the present disclosure includes a tobacco product,prepared from a tobacco plant, or part thereof, produced by growing theseed of tobacco cultivar K326 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13455, where the product is selected from the group consisting of acigarillo, a non-ventilated recess filter cigarette, a vented recessfilter cigarette, a cigar, snuff, and chewing tobacco, and where theproduct has a reduced amount of nornicotine and/or N′-nitrosonornicotine(NNN).

In an aspect, the present disclosure includes a part of a tobacco plant,produced by growing the seed of tobacco cultivar K326 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13455, where the part is selectedfrom the group consisting of leaf, pollen, ovule, embryo, cotyledon,hypocotyl, meristematic cell, protoplast, root, root tip, pistil,anther, flower, shoot, stem, pod and petiole.

In another aspect, the present disclosure includes a tissue cultureproduced from a protoplast or cell from a tobacco plant, or partthereof, produced by growing the seed of tobacco cultivar K326 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13455, where the cell orprotoplast of the tissue culture is produced from a plant part selectedfrom the group consisting of a leaf, pollen, embryo, cotyledon,hypocotyl, meristematic cell, root, root tip, pistil, anther, flower,shoot, stem, pod, and petiole.

In an aspect, the present disclosure includes a tobacco plantregenerated from a tissue culture produced from a protoplast or cellfrom a tobacco plant, or part thereof, produced by growing the seed oftobacco cultivar K326 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-13455,where the cell or protoplast of the tissue culture is produced from aplant part selected from the group consisting of a leaf, pollen, embryo,cotyledon, hypocotyl, meristematic cell, root, root tip, pistil, anther,flower, shoot, stem, pod, and petiole where the regenerated plant hasall, or essentially all of the morphological and physiologicalcharacteristics of cultivar K326 SRC.

In an aspect, the present disclosure includes an F₁ progeny plant oftobacco cultivar K326 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-13455.

In another aspect, the present disclosure includes an F₁ progeny plantof tobacco cultivar K326 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13455, where the F₁ plant is cytoplasmic male sterile (CMS).

In another aspect, the present disclosure includes an F₁ progeny seedproduced by a method of comprising crossing two tobacco plants andharvesting the resultant tobacco seed, where at least one tobacco plantis a tobacco plant produced by growing the seed of tobacco cultivar K326SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13455, where at least onetobacco plant is cytoplasmic male sterile (CMS).

In another aspect, the present disclosure includes a method forproducing a tobacco seed comprising crossing two tobacco plants andharvesting the resultant tobacco seed, where at least one tobacco plantis a tobacco plant produced by growing the seed of tobacco cultivar K326SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13455.

In another aspect, the present disclosure includes a method forproducing a tobacco seed comprising crossing two tobacco plants andharvesting the resultant tobacco seed, where at least one tobacco plantis a tobacco plant produced by growing the seed of tobacco cultivar K326SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13455, where the plant oftobacco cultivar K326 SRC is the male parent.

In another aspect, the present disclosure includes a method forproducing a tobacco seed comprising crossing two tobacco plants andharvesting the resultant tobacco seed, where at least one tobacco plantis a tobacco plant produced by growing the seed of tobacco cultivar K326SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13455, where the plant oftobacco cultivar K326 SRC is the female parent.

In another aspect, the present disclosure includes a method forproducing a tobacco seed comprising crossing two tobacco plants andharvesting the resultant tobacco seed, where at least one tobacco plantis a tobacco plant produced by growing the seed of tobacco cultivar K326SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13455, where at least onetobacco plant is cytoplasmic male sterile (CMS).

In an aspect, the present disclosure also includes a container of F₁progeny seeds produced by a method of comprising crossing two tobaccoplants and harvesting the resultant tobacco seed, where at least onetobacco plant is a tobacco plant produced by growing the seed of tobaccocultivar K326 SRC, a representative sample seed of the cultivar havingbeen deposited with the ATCC under ATCC Accession No. PTA-13455, whereat least one tobacco plant is cytoplasmic male sterile (CMS).

In an aspect, the present disclosure includes an F₁ progeny plantproduced by growing a seed produced by a method of comprising crossingtwo tobacco plants and harvesting the resultant tobacco seed, where atleast one tobacco plant is a tobacco plant produced by growing the seedof tobacco cultivar K326 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13455, and where at least one tobacco plant is cytoplasmic malesterile (CMS).

In an aspect, the present disclosure also includes a harvested leaf ofan F₁ progeny plant produced by growing a seed produced by a method ofcomprising crossing two tobacco plants and harvesting the resultanttobacco seed, where at least one tobacco plant is a tobacco plantproduced by growing the seed of tobacco cultivar K326 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13455, where at least one tobaccoplant is cytoplasmic male sterile (CMS).

In another aspect, the present disclosure further includes a harvestedleaf of an F₁ progeny plant having a reduced amount of nornicotineand/or N′-nitrosonomicotine (NNN) produced by growing a seed produced bya method of comprising crossing two tobacco plants and harvesting theresultant tobacco seed, where at least one tobacco plant is a tobaccoplant produced by growing the seed of tobacco cultivar K326 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13455, where at least one tobaccoplant is cytoplasmic male sterile (CMS).

In an aspect, the present disclosure also includes a harvested leaf ofan F₁ progeny plant having a reduced amount of nornicotine and/orN′-nitrosonornicotine (NNN) in a smoke stream produced from the leaf,where the plant is produced by growing a seed produced by a method ofcomprising crossing two tobacco plants and harvesting the resultanttobacco seed, where at least one tobacco plant is a tobacco plantproduced by growing the seed of tobacco cultivar K326 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13455, where at least one tobaccoplant is cytoplasmic male sterile (CMS).

In an aspect, the present disclosure includes a tobacco product preparedfrom an F₁ progeny tobacco plant, or part thereof, where the plant orpart thereof is produced by growing a F₁ progeny seed produced by amethod comprising crossing two tobacco plants and harvesting theresultant tobacco seed, where at least one tobacco plant is produced bygrowing a seed of tobacco cultivar K326 SRC, a representative sampleseed of the cultivar having been deposited with the ATCC under ATCCAccession No. PTA-13455, where at least one tobacco plant is cytoplasmicmale sterile, and where the tobacco product is selected from the groupconsisting of pipe tobacco, cigar tobacco, cigarette tobacco, chewingtobacco, leaf tobacco, shredded tobacco, and cut tobacco.

In an aspect, the present disclosure further includes a tobacco productprepared from an F₁ progeny tobacco plant, or part thereof, where theplant or part thereof is produced by growing a F₁ progeny seed producedby a method comprising crossing two tobacco plants and harvesting theresultant tobacco seed, where at least one tobacco plant is produced bygrowing a seed of tobacco cultivar K326 SRC, a representative sampleseed of the cultivar having been deposited with the ATCC under ATCCAccession No. PTA-13455, where at least one tobacco plant is cytoplasmicmale sterile, and where the tobacco product is selected from the groupconsisting of a cigarillo, a non-ventilated recess filter cigarette, avented recess filter cigarette, a cigar, snuff, and chewing tobacco.

In an aspect, the present disclosure further includes a tobacco productprepared from an F₁ progeny tobacco plant, or part thereof, where theplant or part thereof is produced by growing a F₁ progeny seed producedby a method comprising crossing two tobacco plants and harvesting theresultant tobacco seed, where at least one tobacco plant is produced bygrowing a seed of tobacco cultivar K326 SRC, a representative sampleseed of the cultivar having been deposited with the ATCC under ATCCAccession No. PTA-13455, where at least one tobacco plant is cytoplasmicmale sterile, and where the tobacco product is selected from the groupconsisting of a cigarillo, a non-ventilated recess filter cigarette, avented recess filter cigarette, a cigar, snuff, and chewing tobacco, andfurther where the product has a reduced amount of nornicotine and/orN′-nitrosonornicotine (NNN).

In another aspect, the present disclosure includes a seed of tobaccocultivar CMS K326 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-13454.

In an aspect, the present disclosure includes a tobacco plant, or partthereof, produced by growing the seed of tobacco cultivar CMS K326 SRC,a representative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13454.

In another aspect, the present disclosure includes a harvested leaf, orpart thereof, of a tobacco plant, or part thereof, produced by growingthe seed of tobacco cultivar CMS K326 SRC, a representative sample seedof the cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-13454.

In an aspect, the present disclosure includes a harvested leaf, or partthereof, of a tobacco plant, produced by growing the seed of tobaccocultivar CMS K326 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-13454,where the leaf has a reduced amount of nornicotine and/orN′-nitrosonornicotine (NNN).

In another aspect, the present disclosure includes a harvested leaf, orpart thereof, produced by growing the seed of tobacco cultivar CMS K326SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13454, where the leaf has areduced amount of nornicotine and/or N′-nitrosonornicotine (NNN), wherethe reduced amount of nornicotine and/or N′-nitrosonomicotine (NNN) isreduced in a smoke stream produced from the leaf.

In a further aspect, the present disclosure includes a tobacco product,prepared from a tobacco plant, or part thereof, produced by growing theseed of tobacco cultivar CMS K326 SRC, a representative sample seed ofthe cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-13454, where the product is selected from the group consistingof pipe tobacco, cigar tobacco, cigarette tobacco, chewing tobacco, leaftobacco, shredded tobacco, and cut tobacco.

In an aspect, the present disclosure includes a tobacco product,prepared from a tobacco plant, or part thereof, produced by growing theseed of tobacco cultivar CMS K326 SRC, a representative sample seed ofthe cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-13454, where the product is selected from the group consistingof a cigarillo, a non-ventilated recess filter cigarette, a ventedrecess filter cigarette, a cigar, snuff, and chewing tobacco.

In another aspect, the present disclosure includes a tobacco product,prepared from a tobacco plant, or part thereof, produced by growing theseed of tobacco cultivar CMS K326 SRC, a representative sample seed ofthe cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-13454, where the product is selected from the group consistingof a cigarillo, a non-ventilated recess filter cigarette, a ventedrecess filter cigarette, a cigar, snuff, and chewing tobacco where theproduct has a reduced amount of nornicotine and/or N′-nitrosonornicotine(NNN).

In an aspect, the present disclosure includes a part of a tobacco plant,produced by growing the seed of tobacco cultivar CMS K326 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13454, where the part is selectedfrom the group consisting of leaf, pollen, ovule, embryo, cotyledon,hypocotyl, meristematic cell, protoplast, root, root tip, pistil,anther, flower, shoot, stem, pod, and petiole.

In another aspect, the present disclosure includes a tissue cultureproduced from a protoplast or cell from a tobacco plant, or partthereof, produced by growing the seed of tobacco cultivar CMS K326 SRC,a representative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13454, where the cell orprotoplast of the tissue culture is produced from a plant part selectedfrom the group consisting of a leaf, pollen, embryo, cotyledon,hypocotyl, meristematic cell, root, root tip, pistil, anther, flower,shoot, stem, pod, and petiole.

In an aspect, the present disclosure includes a tobacco plantregenerated from a tissue culture produced from a protoplast or cellfrom a tobacco plant, or part thereof, produced by growing the seed oftobacco cultivar CMS K326 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13454, where the cell or protoplast of the tissue culture isproduced from a plant part selected from the group consisting of a leaf,pollen, embryo, cotyledon, hypocotyl, meristematic cell, root, root tip,pistil, anther, flower, shoot, stem, pod, and petiole where theregenerated plant has all, or essentially all of the morphological andphysiological characteristics of cultivar CMS K326 SRC.

In an aspect, the present disclosure includes an F₁ progeny plant oftobacco cultivar CMS K326 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13454.

In another aspect, the present disclosure includes an F₁ progeny plantof tobacco cultivar CMS K326 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13454, where the F₁ plant is cytoplasmic male sterile (CMS).

In another aspect, the present disclosure includes an F₁ progeny seedproduced by a method of comprising crossing two tobacco plants andharvesting the resultant tobacco seed, where at least one tobacco plantis cytoplasmic male sterile (CMS), where the cytoplasmic male sterileplant is a plant of tobacco cultivar CMS K326 SRC, a representativesample seed of the cultivar having been deposited with the ATCC underATCC Accession No. PTA-13454.

In another aspect, the present disclosure includes a method forproducing a tobacco seed comprising crossing two tobacco plants andharvesting the resultant tobacco seed, where at least one tobacco plantis a tobacco plant produced by growing the seed of tobacco cultivar CMSK326 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No. PTA-13454.

In another aspect, the present disclosure includes a method forproducing a tobacco seed comprising crossing two tobacco plants andharvesting the resultant tobacco seed, where at least one tobacco plantis a tobacco plant produced by growing the seed of tobacco cultivar CMSK326 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No, PTA-13454, where theplant of tobacco cultivar CMS K326 SRC is the female parent.

In another aspect, the present disclosure includes a method forproducing a tobacco seed comprising crossing two tobacco plants andharvesting the resultant tobacco seed, where at least one tobacco plantis cytoplasmic male sterile (CMS), where the cytoplasmic male sterileplant is a plant of tobacco cultivar CMS K326 SRC, a representativesample seed of the cultivar having been deposited with the ATCC underATCC Accession No. PTA-13454.

In an aspect, the present disclosure also includes a container of F₁progeny seeds produced by a method of comprising crossing two tobaccoplants and harvesting the resultant tobacco seed, where at least onetobacco plant is cytoplasmic male sterile (CMS), where the cytoplasmicmale sterile plant is a tobacco plant produced by growing the seed oftobacco cultivar CMS K326 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13454.

In an aspect, the present disclosure includes an F₁ progeny plantproduced by growing a seed produced by a method of comprising crossingtwo tobacco plants and harvesting the resultant tobacco seed, where atleast one tobacco plant is cytoplasmic male sterile (CMS), where thecytoplasmic male sterile plant is a tobacco plant produced by growingthe seed of tobacco cultivar CMS K326 SRC, a representative sample seedof the cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-13454.

In an aspect, the present disclosure also includes a harvested leaf ofan F₁ progeny plant produced by growing a seed produced by a method ofcomprising crossing two tobacco plants and harvesting the resultanttobacco seed, where at least one tobacco plant is cytoplasmic malesterile (CMS), where the cytoplasmic male sterile plant is a tobaccoplant produced by growing the seed of tobacco cultivar CMS K326 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13454.

In another aspect, the present disclosure further includes a harvestedleaf of an F₁ progeny plant having a reduced amount of nornicotineand/or N′-nitrosonomicotine (NNN) produced by growing a seed produced bya method of comprising crossing two tobacco plants and harvesting theresultant tobacco seed, where at least one tobacco plant is cytoplasmicmale sterile (CMS), where the cytoplasmic male sterile plant is atobacco plant produced by growing the seed of tobacco cultivar CMS K326SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13454.

In an aspect, the present disclosure also includes a harvested leaf ofan F₁ progeny plant having a reduced amount of nornicotine and/orN′-nitrosonornicotine (NNN) in a smoke stream produced from the leaf,where the plant is produced by growing a seed produced by a method ofcomprising crossing two tobacco plants and harvesting the resultanttobacco seed, where at least one tobacco plant is cytoplasmic malesterile (CMS), where the cytoplasmic male sterile plant is a tobaccoplant produced by growing the seed of tobacco cultivar CMS K326 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13454.

In an aspect, the present disclosure includes a tobacco product preparedfrom an F₁ progeny tobacco plant, or part thereof, where the plant orpart thereof is produced by growing a F₁ progeny seed produced by amethod comprising crossing two tobacco plants and harvesting theresultant tobacco seed, where at least one tobacco plant is cytoplasmicmale sterile (CMS), where the cytoplasmic male sterile plant is atobacco plant produced by growing the seed of tobacco cultivar CMS K326SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13454, and where the tobaccoproduct is selected from the group consisting of pipe tobacco, cigartobacco, cigarette tobacco, chewing tobacco, leaf tobacco, shreddedtobacco, and cut tobacco.

In an aspect, the present disclosure further includes a tobacco productprepared from an F₁ progeny tobacco plant, or part thereof, where theplant or part thereof is produced by growing a F₁ progeny seed producedby a method comprising crossing two tobacco plants and harvesting theresultant tobacco seed, where at least one tobacco plant is cytoplasmicmale sterile (CMS), where the cytoplasmic male sterile plant is atobacco plant produced by growing the seed of tobacco cultivar CMS K326SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13454, and where the tobaccoproduct is selected from the group consisting of a cigarillo, anon-ventilated recess filter cigarette, a vented recess filtercigarette, a cigar, snuff, and chewing tobacco.

In an aspect, the present disclosure further includes a tobacco productprepared from an F₁ progeny tobacco plant, or part thereof, where theplant or part thereof is produced by growing a F₁ progeny seed producedby a method comprising crossing two tobacco plants and harvesting theresultant tobacco seed, where at least one tobacco plant is cytoplasmicmale sterile (CMS), where the cytoplasmic male sterile plant is atobacco plant produced by growing the seed of tobacco cultivar CMS K326SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13454, and where the tobaccoproduct is selected from the group consisting of a cigarillo, anon-ventilated recess filter cigarette, a vented recess filtercigarette, a cigar, snuff, and chewing tobacco, and further where theproduct has a reduced amount of nornicotine and/or N′-nitrosonomicotine(NNN).

In an aspect, the present disclosure includes a method for producing atobacco seed comprising crossing two tobacco plants and harvesting theresultant tobacco seed, where at least one tobacco plant is cytoplasmicmale sterile, where the cytoplasmic male sterile plant is a plant oftobacco cultivar CMS K326 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13454.

In another aspect, the present disclosure includes a seed of tobaccocultivar K346 SRC, a representative sample seed of the cultivar havingbeen deposited with the ATCC under ATCC Accession No. PTA-13453.

In an aspect, the present disclosure includes a tobacco plant, or partthereof, produced by growing the seed of tobacco cultivar K346 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13453.

In another aspect, the present disclosure includes a harvested leaf, orpart thereof, of a tobacco plant, or part thereof, produced by growingthe seed of tobacco cultivar K346 SRC, a representative sample seed ofthe cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-13453.

In an aspect, the present disclosure includes a harvested leaf, or partthereof, of a tobacco plant, produced by growing the seed of tobaccocultivar K346 SRC, a representative sample seed of the cultivar havingbeen deposited with the ATCC under ATCC Accession No. PTA-13453, wherethe leaf has a reduced amount of nornicotine and/orN′-nitrosonornicotine (NNN).

In another aspect, the present disclosure includes a harvested leaf, orpart thereof, produced by growing the seed of tobacco cultivar K346 SRC,a representative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13453, where the leaf has areduced amount of nornicotine and/or N′-nitrosonornicotine (NNN), wherethe reduced amount of nornicotine and/or N′-nitrosonornicotine (NNN) isreduced in a smoke stream produced from the leaf.

In a further aspect, the present disclosure includes a tobacco product,prepared from a tobacco plant, or part thereof, produced by growing theseed of tobacco cultivar K346 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13453, where the product is selected from the group consisting ofpipe tobacco, cigar tobacco, cigarette tobacco, chewing tobacco, leaftobacco, shredded tobacco, and cut tobacco.

In an aspect, the present disclosure includes a tobacco product,prepared from a tobacco plant, or part thereof, produced by growing theseed of tobacco cultivar K346 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13453, where the product is selected from the group consisting of acigarillo, a non-ventilated recess filter cigarette, a vented recessfilter cigarette, a cigar, snuff, and chewing tobacco.

In another aspect, the present disclosure includes a tobacco product,prepared from a tobacco plant, or part thereof, produced by growing theseed of tobacco cultivar K346 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13453, where the product is selected from the group consisting of acigarillo, a non-ventilated recess filter cigarette, a vented recessfilter cigarette, a cigar, snuff, and chewing tobacco where the producthas a reduced amount of nornicotine and/or N′-nitrosonornicotine (NNN).

In an aspect, the present disclosure includes a part of a tobacco plant,produced by growing the seed of tobacco cultivar K346 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13453, where the part is selectedfrom the group consisting of leaf, pollen, ovule, embryo, cotyledon,hypocotyl, meristematic cell, protoplast, root, root tip, pistil,anther, flower, shoot, stem, pod, and petiole.

In another aspect, the present disclosure includes a tissue cultureproduced from a protoplast or cell from a tobacco plant, or partthereof, produced by growing the seed of tobacco cultivar K346 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13453, where the cell orprotoplast of the tissue culture is produced from a plant part selectedfrom the group consisting of a leaf, pollen, embryo, cotyledon,hypocotyl, meristematic cell, root, root tip, pistil, anther, flower,shoot, stem, pod, and petiole.

In an aspect, the present disclosure includes a tobacco plantregenerated from a tissue culture produced from a protoplast or cellfrom a tobacco plant, or part thereof, produced by growing the seed oftobacco cultivar K346 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-13453,where the cell or protoplast of the tissue culture is produced from aplant part selected from the group consisting of a leaf, pollen, embryo,cotyledon, hypocotyl, meristematic cell, root, root tip, pistil, anther,flower, shoot, stem, pod, and petiole where the regenerated plant hasall, or essentially all of the morphological and physiologicalcharacteristics of cultivar K346 SRC.

In an aspect, the present disclosure includes an F₁ progeny plant oftobacco cultivar K346 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-13453.

In another aspect, the present disclosure includes an F₁ progeny plantof tobacco cultivar K346 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13453, where the F₁ plant is cytoplasmic male sterile (CMS).

In another aspect, the present disclosure includes a method forproducing a tobacco seed comprising crossing two tobacco plants andharvesting the resultant tobacco seed, where at least one tobacco plantis a tobacco plant produced by growing the seed of tobacco cultivar K346SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13453.

In another aspect, the present disclosure includes a method forproducing a tobacco seed comprising crossing two tobacco plants andharvesting the resultant tobacco seed, where at least one tobacco plantis a tobacco plant produced by growing the seed of tobacco cultivar K346SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13453, where the plant oftobacco cultivar K346 SRC is the male parent.

In another aspect, the present disclosure includes a method forproducing a tobacco seed comprising crossing two tobacco plants andharvesting the resultant tobacco seed, where at least one tobacco plantis a tobacco plant produced by growing the seed of tobacco cultivar K346SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13453, where the plant oftobacco cultivar K346 SRC is the female parent.

In another aspect, the present disclosure includes a method forproducing a tobacco seed comprising crossing two tobacco plants andharvesting the resultant tobacco seed, where at least one tobacco plantis a tobacco plant produced by growing the seed of tobacco cultivar K346SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13453, where at least onetobacco plant is cytoplasmic male sterile (CMS).

In another aspect, the present disclosure includes an F₁ progeny seedproduced by a method of comprising crossing two tobacco plants andharvesting the resultant tobacco seed, where at least one tobacco plantis a tobacco plant produced by growing the seed of tobacco cultivar K346SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13453, where at least onetobacco plant is cytoplasmic male sterile (CMS).

In an aspect, the present disclosure also includes a container of F₁progeny seeds produced by a method of comprising crossing two tobaccoplants and harvesting the resultant tobacco seed, where at least onetobacco plant is a tobacco plant produced by growing the seed of tobaccocultivar K346 SRC, a representative sample seed of the cultivar havingbeen deposited with the ATCC under ATCC Accession No. PTA-13453, whereat least one tobacco plant is cytoplasmic male sterile (CMS).

In an aspect, the present disclosure includes an F₁ progeny plantproduced by growing a seed produced by a method of comprising crossingtwo tobacco plants and harvesting the resultant tobacco seed, where atleast one tobacco plant is a tobacco plant produced by growing the seedof tobacco cultivar K346 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13453, and where at least one tobacco plant is cytoplasmic malesterile (CMS).

In an aspect, the present disclosure also includes a harvested leaf ofan F₁ progeny plant produced by growing a seed produced by a method ofcomprising crossing two tobacco plants and harvesting the resultanttobacco seed, where at least one tobacco plant is a tobacco plantproduced by growing the seed of tobacco cultivar K346 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13453, where at least one tobaccoplant is cytoplasmic male sterile (CMS).

In another aspect, the present disclosure further includes a harvestedleaf of an F₁ progeny plant having a reduced amount of nornicotineand/or N′-nitrosonornicotine (NNN) produced by growing a seed producedby a method of comprising crossing two tobacco plants and harvesting theresultant tobacco seed, where at least one tobacco plant is a tobaccoplant produced by growing the seed of tobacco cultivar K346 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13453, where at least one tobaccoplant is cytoplasmic male sterile (CMS).

In an aspect, the present disclosure also includes a harvested leaf ofan F₁ progeny plant having a reduced amount of nornicotine and/orN′-nitrosonornicotine (NNN) in a smoke stream produced from the leaf,where the plant is produced by growing a seed produced by a method ofcomprising crossing two tobacco plants and harvesting the resultanttobacco seed, where at least one tobacco plant is a tobacco plantproduced by growing the seed of tobacco cultivar K346 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13453, where at least one tobaccoplant is cytoplasmic male sterile (CMS).

In an aspect, the present disclosure includes a tobacco product preparedfrom an F₁ progeny tobacco plant, or part thereof, where the plant orpart thereof is produced by growing a F₁ progeny seed produced by amethod comprising crossing two tobacco plants and harvesting theresultant tobacco seed, where at least one tobacco plant is produced bygrowing a seed of tobacco cultivar K346 SRC, a representative sampleseed of the cultivar having been deposited with the ATCC under ATCCAccession No. PTA-13453, where at least one tobacco plant is cytoplasmicmale sterile, and where the tobacco product is selected from the groupconsisting of pipe tobacco, cigar tobacco, cigarette tobacco, chewingtobacco, leaf tobacco, shredded tobacco, and cut tobacco.

In an aspect, the present disclosure further includes a tobacco productprepared from an F₁ progeny tobacco plant, or part thereof, where theplant or part thereof is produced by growing a F₁ progeny seed producedby a method comprising crossing two tobacco plants and harvesting theresultant tobacco seed, where at least one tobacco plant is produced bygrowing a seed of tobacco cultivar K346 SRC, a representative sampleseed of the cultivar having been deposited with the ATCC under ATCCAccession No. PTA-13453, where at least one tobacco plant is cytoplasmicmale sterile, and where the tobacco product is selected from the groupconsisting of a cigarillo, a non-ventilated recess filter cigarette, avented recess filter cigarette, a cigar, snuff, and chewing tobacco.

In an aspect, the present disclosure further includes a tobacco productprepared from an F₁ progeny tobacco plant, or part thereof, where theplant or part thereof is produced by growing a F₁ progeny seed producedby a method comprising crossing two tobacco plants and harvesting theresultant tobacco seed, where at least one tobacco plant is produced bygrowing a seed of tobacco cultivar K346 SRC, a representative sampleseed of the cultivar having been deposited with the ATCC under ATCCAccession No. PTA-13453, where at least one tobacco plant is cytoplasmicmale sterile, and where the tobacco product is selected from the groupconsisting of a cigarillo, a non-ventilated recess filter cigarette, avented recess filter cigarette, a cigar, snuff, and chewing tobacco, andfurther where the product has a reduced amount of nornicotine and/orN′-nitrosonornicotine (NNN).

In another aspect, the present disclosure includes a seed of tobaccocultivar CMS K346 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-13452.

In an aspect, the present disclosure includes a tobacco plant, or partthereof, produced by growing the seed of tobacco cultivar CMS K346 SRC,a representative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13452.

In another aspect, the present disclosure includes a harvested leaf, orpart thereof, of a tobacco plant, or part thereof, produced by growingthe seed of tobacco cultivar CMS K346 SRC, a representative sample seedof the cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-13452.

In an aspect, the present disclosure includes a harvested leaf, or partthereof, of a tobacco plant, produced by growing the seed of tobaccocultivar CMS K346 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-13452,where the leaf has a reduced amount of nornicotine and/orN′-nitrosonornicotine (NNN).

In another aspect, the present disclosure includes a harvested leaf, orpart thereof, produced by growing the seed of tobacco cultivar CMS K346SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13452, where the leaf has areduced amount of nornicotine and/or N′-nitrosonomicotine (NNN), wherethe reduced amount of nornicotine and/or N′-nitrosonornicotine (NNN) isreduced in a smoke stream produced from the leaf.

In a further aspect, the present disclosure includes a tobacco product,prepared from a tobacco plant, or part thereof, produced by growing theseed of tobacco cultivar CMS K346 SRC, a representative sample seed ofthe cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-13452, where the product is selected from the group consistingof pipe tobacco, cigar tobacco, cigarette tobacco, chewing tobacco, leaftobacco, shredded tobacco, and cut tobacco.

In an aspect, the present disclosure includes a tobacco product,prepared from a tobacco plant, or part thereof, produced by growing theseed of tobacco cultivar CMS K346 SRC, a representative sample seed ofthe cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-13452, where the product is selected from the group consistingof a cigarillo, a non-ventilated recess filter cigarette, a ventedrecess filter cigarette, a cigar, snuff, and chewing tobacco.

In another aspect, the present disclosure includes a tobacco product,prepared from a tobacco plant, or part thereof, produced by growing theseed of tobacco cultivar CMS K346 SRC, a representative sample seed ofthe cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-13452, where the product is selected from the group consistingof a cigarillo, a non-ventilated recess filter cigarette, a ventedrecess filter cigarette, a cigar, snuff, and chewing tobacco where theproduct has a reduced amount of nornicotine and/or N′-nitrosonornicotine(NNN).

In an aspect, the present disclosure includes a part of a tobacco plant,produced by growing the seed of tobacco cultivar CMS K346 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13452, where the part is selectedfrom the group consisting of leaf, pollen, ovule, embryo, cotyledon,hypocotyl, meristematic cell, protoplast, root, root tip, pistil,anther, flower, shoot, stem, pod, and petiole.

In another aspect, the present disclosure includes a tissue cultureproduced from a protoplast or cell from a tobacco plant, or partthereof, produced by growing the seed of tobacco cultivar CMS K346 SRC,a representative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13452, where the cell orprotoplast of the tissue culture is produced from a plant part selectedfrom the group consisting of a leaf, pollen, embryo, cotyledon,hypocotyl, meristematic cell, root, root tip, pistil, anther, flower,shoot, stem, pod, and petiole.

In an aspect, the present disclosure includes a tobacco plantregenerated from a tissue culture produced from a protoplast or cellfrom a tobacco plant, or part thereof, produced by growing the seed oftobacco cultivar CMS K346 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13452, where the cell or protoplast of the tissue culture isproduced from a plant part selected from the group consisting of a leaf,pollen, embryo, cotyledon, hypocotyl, meristematic cell, root, root tip,pistil, anther, flower, shoot, stem, pod, and petiole where theregenerated plant has all, or essentially all of the morphological andphysiological characteristics of cultivar CMS K346 SRC.

In an aspect, the present disclosure includes an F₁ progeny plant oftobacco cultivar CMS K346 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC AccessionPTA-13452.

In another aspect, the present disclosure includes an F₁ progeny plantof tobacco cultivar CMS K346 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13452, where the F₁ plant is cytoplasmic male sterile (CMS).

In another aspect, the present disclosure includes an F₁ progeny seedproduced by a method of comprising crossing two tobacco plants andharvesting the resultant tobacco seed, where at least one tobacco plantis cytoplasmic male sterile (CMS), where the cytoplasmic male sterileplant is a plant of tobacco cultivar CMS K346 SRC, a representativesample seed of the cultivar having been deposited with the ATCC underATCC Accession No. PTA-13452.

In another aspect, the present disclosure includes a method forproducing a tobacco seed comprising crossing two tobacco plants andharvesting the resultant tobacco seed, where at least one tobacco plantis a tobacco plant produced by growing the seed of tobacco cultivar CMSK346 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No. PTA-13452.

In another aspect, the present disclosure includes a method forproducing a tobacco seed comprising crossing two tobacco plants andharvesting the resultant tobacco seed, where at least one tobacco plantis a tobacco plant produced by growing the seed of tobacco cultivar CMSK346 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No, PTA-13452, where theplant of tobacco cultivar CMS K346 SRC is the female parent.

In an aspect, the present disclosure includes a method for producing atobacco seed comprising crossing two tobacco plants and harvesting theresultant tobacco seed, where at least one tobacco plant is cytoplasmicmale sterile, where the cytoplasmic male sterile plant is a plant oftobacco cultivar CMS K346 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13452.

In an aspect, the present disclosure also includes a container of F₁progeny seeds produced by a method of comprising crossing two tobaccoplants and harvesting the resultant tobacco seed, where at least onetobacco plant is cytoplasmic male sterile (CMS), where the cytoplasmicmale sterile plant is a tobacco plant produced by growing the seed oftobacco cultivar CMS K346 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13452.

In an aspect, the present disclosure includes an F₁ progeny plantproduced by growing a seed produced by a method of comprising crossingtwo tobacco plants and harvesting the resultant tobacco seed, where atleast one tobacco plant is cytoplasmic male sterile (CMS), where thecytoplasmic male sterile plant is a tobacco plant produced by growingthe seed of tobacco cultivar CMS K346 SRC, a representative sample seedof the cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-13452.

In an aspect, the present disclosure also includes a harvested leaf ofan F₁ progeny plant produced by growing a seed produced by a method ofcomprising crossing two tobacco plants and harvesting the resultanttobacco seed, where at least one tobacco plant is cytoplasmic malesterile (CMS), where the cytoplasmic male sterile plant is a tobaccoplant produced by growing the seed of tobacco cultivar CMS K346 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13452.

In another aspect, the present disclosure further includes a harvestedleaf of an F₁ progeny plant having a reduced amount of nornicotineand/or N′-nitrosonornicotine (NNN) produced by growing a seed producedby a method of comprising crossing two tobacco plants and harvesting theresultant tobacco seed, where at least one tobacco plant is cytoplasmicmale sterile (CMS), where the cytoplasmic male sterile plant is atobacco plant produced by growing the seed of tobacco cultivar CMS K346SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13452.

In an aspect, the present disclosure also includes a harvested leaf ofan F₁ progeny plant having a reduced amount of nornicotine and/orN′-nitrosonornicotine (NNN) in a smoke stream produced from the leaf,where the plant is produced by growing a seed produced by a method ofcomprising crossing two tobacco plants and harvesting the resultanttobacco seed, where at least one tobacco plant is cytoplasmic malesterile (CMS), where the cytoplasmic male sterile plant is a tobaccoplant produced by growing the seed of tobacco cultivar CMS K346 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13452.

In an aspect, the present disclosure includes a tobacco product preparedfrom an F₁ progeny tobacco plant, or part thereof, where the plant orpart thereof is produced by growing a F₁ progeny seed produced by amethod comprising crossing two tobacco plants and harvesting theresultant tobacco seed, where at least one tobacco plant is cytoplasmicmale sterile (CMS), where the cytoplasmic male sterile plant is atobacco plant produced by growing the seed of tobacco cultivar CMS K346SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13452, and where the tobaccoproduct is selected from the group consisting of pipe tobacco, cigartobacco, cigarette tobacco, chewing tobacco, leaf tobacco, shreddedtobacco, and cut tobacco.

In an aspect, the present disclosure further includes a tobacco productprepared from an F₁ progeny tobacco plant, or part thereof, where theplant or part thereof is produced by growing a F₁ progeny seed producedby a method comprising crossing two tobacco plants and harvesting theresultant tobacco seed, where at least one tobacco plant is cytoplasmicmale sterile (CMS), where the cytoplasmic male sterile plant is atobacco plant produced by growing the seed of tobacco cultivar CMS K346SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13452, and where the tobaccoproduct is selected from the group consisting of a cigarillo, anon-ventilated recess filter cigarette, a vented recess filtercigarette, a cigar, snuff, and chewing tobacco.

In an aspect, the present disclosure further includes a tobacco productprepared from an F₁ progeny tobacco plant, or part thereof, where theplant or part thereof is produced by growing a F₁ progeny seed producedby a method comprising crossing two tobacco plants and harvesting theresultant tobacco seed, where at least one tobacco plant is cytoplasmicmale sterile (CMS), where the cytoplasmic male sterile plant is atobacco plant produced by growing the seed of tobacco cultivar CMS K346SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13452, and where the tobaccoproduct is selected from the group consisting of a cigarillo, anon-ventilated recess filter cigarette, a vented recess filtercigarette, a cigar, snuff, and chewing tobacco, and further where theproduct has a reduced amount of nornicotine and/or N′-nitrosonornicotine(NNN).

In an aspect, the present disclosure includes a seed of tobacco cultivarNC1562-1 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No. PTA-13457.

In another aspect, the present disclosure includes a tobacco plant, orpart thereof, produced by growing the seed of tobacco cultivar NC1562-1SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13457.

In a further aspect, the present disclosure includes a harvested leaf,or part thereof, of a tobacco plant, or part thereof, produced bygrowing the seed of tobacco cultivar NC1562-1 SRC, a representativesample seed of the cultivar having been deposited with the ATCC underATCC Accession No. PTA-13457.

In an aspect, the present disclosure includes a harvested leaf, or partthereof, of a tobacco plant, produced by growing the seed of tobaccocultivar NC1562-1 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-13457,where the leaf has a reduced amount of nornicotine and/orN′-nitrosonomicotine (NNN).

In an aspect, the present disclosure includes a harvested leaf, or partthereof, produced by growing the seed of tobacco cultivar NC1562-1 SRC,a representative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13457, where the leaf has areduced amount of nornicotine and/or N′-nitrosonornicotine (NNN), wherethe reduced amount of nornicotine and/or N′-nitrosonornicotine (NNN) isreduced in a smoke stream produced from the leaf.

In a further aspect, the present disclosure includes a tobacco product,prepared from a tobacco plant, or part thereof, produced by growing theseed of tobacco cultivar NC1562-1 SRC, a representative sample seed ofthe cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-13457, where the product is selected from the group consistingof pipe tobacco, cigar tobacco, cigarette tobacco, chewing tobacco, leaftobacco, shredded tobacco, and cut tobacco.

In an aspect, the present disclosure includes a tobacco product,prepared from a tobacco plant, or part thereof, produced by growing theseed of tobacco cultivar NC1562-1 SRC, a representative sample seed ofthe cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-13457, where the product is selected from the group consistingof a cigarillo, a non-ventilated recess filter cigarette, a ventedrecess filter cigarette, a cigar, snuff, and chewing tobacco.

In another aspect, the present disclosure includes a tobacco product,prepared from a tobacco plant, or part thereof, produced by growing theseed of tobacco cultivar NC1562-1 SRC, a representative sample seed ofthe cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-13457, where the product is selected from the group consistingof a cigarillo, a non-ventilated recess filter cigarette, a ventedrecess filter cigarette, a cigar, snuff, and chewing tobacco, and wherethe product has a reduced amount of nornicotine and/orN′-nitrosonornicotine (NNN).

In an aspect, the present disclosure includes a part of a tobacco plant,produced by growing the seed of tobacco cultivar NC1562-1 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13457, where the part is selectedfrom the group consisting of leaf, pollen, ovule, embryo, cotyledon,hypocotyl, meristematic cell, protoplast, root, root tip, pistil,anther, flower, shoot, stem, pod, and petiole.

In another aspect, the present disclosure includes a tissue cultureproduced from a protoplast or cell from a tobacco plant, or partthereof, produced by growing the seed of tobacco cultivar NC1562-1 SRC,a representative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13457, where the cell orprotoplast of the tissue culture is produced from a plant part selectedfrom the group consisting of a leaf, pollen, embryo, cotyledon,hypocotyl, meristematic cell, root, root tip, pistil, anther, flower,shoot, stem, pod, and petiole.

In an aspect, the present disclosure includes a tobacco plantregenerated from a tissue culture produced from a protoplast or cellfrom a tobacco plant, or part thereof, produced by growing the seed oftobacco cultivar NC1562-1 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13457, where the cell or protoplast of the tissue culture isproduced from a plant part selected from the group consisting of a leaf,pollen, embryo, cotyledon, hypocotyl, meristematic cell, root, root tip,pistil, anther, flower, shoot, stem, pod, and petiole where theregenerated plant has all, or essentially all of the morphological andphysiological characteristics of cultivar NC1562-1 SRC.

In an aspect, the present disclosure includes an F₁ progeny plant oftobacco cultivar NC1562-1 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13457.

In another aspect, the present disclosure includes an F₁ progeny plantof tobacco cultivar NC1562-1 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13457, where the F₁ plant is cytoplasmic male sterile (CMS).

In another aspect, the present disclosure includes an F₁ progeny seedproduced by a method of comprising crossing two tobacco plants andharvesting the resultant tobacco seed, where at least one tobacco plantis a tobacco plant produced by growing the seed of tobacco cultivarNC1562-1 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No. PTA-13457, where atleast one tobacco plant is cytoplasmic male sterile (CMS).

In another aspect, the present disclosure includes a method forproducing a tobacco seed comprising crossing two tobacco plants andharvesting the resultant tobacco seed, where at least one tobacco plantis a tobacco plant produced by growing the seed of tobacco cultivarNC1562-1 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No. PTA-13457.

In another aspect, the present disclosure includes a method forproducing a tobacco seed comprising crossing two tobacco plants andharvesting the resultant tobacco seed, where at least one tobacco plantis a tobacco plant produced by growing the seed of tobacco cultivarNC1562-1 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No. PTA-13457, where theplant of tobacco cultivar NC1562-1 SRC is the female parent.

In another aspect, the present disclosure includes a method forproducing a tobacco seed comprising crossing two tobacco plants andharvesting the resultant tobacco seed, where at least one tobacco plantis a tobacco plant produced by growing the seed of tobacco cultivarNC1562-1 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No. PTA-13457, where theplant of tobacco cultivar NC1562-1 SRC is the male parent.

In another aspect, the present disclosure includes a method forproducing a tobacco seed comprising crossing two tobacco plants andharvesting the resultant tobacco seed, where at least one tobacco plantis a tobacco plant produced by growing the seed of tobacco cultivarNC1562-1 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No. PTA-13457, where atleast one tobacco plant is cytoplasmic male sterile (CMS).

In an aspect, the present disclosure also includes a container of F₁progeny seeds produced by a method of comprising crossing two tobaccoplants and harvesting the resultant tobacco seed, where at least onetobacco plant is a tobacco plant produced by growing the seed of tobaccocultivar NC1562-1 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-13457,where at least one tobacco plant is cytoplasmic male sterile (CMS).

In an aspect, the present disclosure includes an F₁ progeny plantproduced by growing a seed produced by a method of comprising crossingtwo tobacco plants and harvesting the resultant tobacco seed, where atleast one tobacco plant is a tobacco plant produced by growing the seedof tobacco cultivar NC1562-1 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13457, and where at least one tobacco plant is cytoplasmic malesterile (CMS).

In an aspect, the present disclosure also includes a harvested leaf ofan F₁ progeny plant produced by growing a seed produced by a method ofcomprising crossing two tobacco plants and harvesting the resultanttobacco seed, where at least one tobacco plant is a tobacco plantproduced by growing the seed of tobacco cultivar NC1562-1 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13457, where at least one tobaccoplant is cytoplasmic male sterile (CMS).

In another aspect, the present disclosure further includes a harvestedleaf of an F₁ progeny plant having a reduced amount of nornicotineand/or N′-nitrosonornicotine (NNN) produced by growing a seed producedby a method of comprising crossing two tobacco plants and harvesting theresultant tobacco seed, where at least one tobacco plant is a tobaccoplant produced by growing the seed of tobacco cultivar NC1562-1 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13457, where at least one tobaccoplant is cytoplasmic male sterile (CMS).

In an aspect, the present disclosure also includes a harvested leaf ofan F₁ progeny plant having a reduced amount of nornicotine and/orN′-nitrosonomicotine (NNN) in a smoke stream produced from the leaf,where the plant is produced by growing a seed produced by a method ofcomprising crossing two tobacco plants and harvesting the resultanttobacco seed, where at least one tobacco plant is a tobacco plantproduced by growing the seed of tobacco cultivar NC1562-1 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13457, where at least one tobaccoplant is cytoplasmic male sterile (CMS).

In an aspect, the present disclosure includes a tobacco product preparedfrom an F₁ progeny tobacco plant, or part thereof, where the plant orpart thereof is produced by growing a F₁ progeny seed produced by amethod comprising crossing two tobacco plants and harvesting theresultant tobacco seed, where at least one tobacco plant is produced bygrowing a seed of tobacco cultivar NC1562-1 SRC, a representative sampleseed of the cultivar having been deposited with the ATCC under ATCCAccession No. PTA-13457, where at least one tobacco plant is cytoplasmicmale sterile, and where the tobacco product is selected from the groupconsisting of pipe tobacco, cigar tobacco, cigarette tobacco, chewingtobacco, leaf tobacco, shredded tobacco, and cut tobacco.

In an aspect, the present disclosure further includes a tobacco productprepared from an F₁ progeny tobacco plant, or part thereof, where theplant or part thereof is produced by growing a F₁ progeny seed producedby a method comprising crossing two tobacco plants and harvesting theresultant tobacco seed, where at least one tobacco plant is produced bygrowing a seed of tobacco cultivar NC1562-1 SRC, a representative sampleseed of the cultivar having been deposited with the ATCC under ATCCAccession No. PTA-13457, where at least one tobacco plant is cytoplasmicmale sterile, and where the tobacco product is selected from the groupconsisting of a cigarillo, a non-ventilated recess filter cigarette, avented recess filter cigarette, a cigar, snuff, and chewing tobacco.

In an aspect, the present disclosure further includes a tobacco productprepared from an F₁ progeny tobacco plant, or part thereof, where theplant or part thereof is produced by growing a F₁ progeny seed producedby a method comprising crossing two tobacco plants and harvesting theresultant tobacco seed, where at least one tobacco plant is produced bygrowing a seed of tobacco cultivar NC1562-1 SRC, a representative sampleseed of the cultivar having been deposited with the ATCC under ATCCAccession No. PTA-13457, where at least one tobacco plant is cytoplasmicmale sterile, and where the tobacco product is selected from the groupconsisting of a cigarillo, a non-ventilated recess filter cigarette, avented recess filter cigarette, a cigar, snuff, and chewing tobacco, andfurther where the product has a reduced amount of nornicotine and/orN′-nitrosonornicotine (NNN).

In an aspect, the present disclosure includes a seed of tobacco cultivarNCTG-61 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No. PTA-120314.

In another aspect, the present disclosure includes a tobacco plant, orpart thereof, produced by growing the seed of tobacco cultivar NCTG-61SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-120314.

In a further aspect, the present disclosure includes a harvested leaf,or part thereof, of a tobacco plant, or part thereof, produced bygrowing the seed of tobacco cultivar NCTG-61 SRC, a representativesample seed of the cultivar having been deposited with the ATCC underATCC Accession No. PTA-120314.

In an aspect, the present disclosure includes a harvested leaf, or partthereof, of a tobacco plant, produced by growing the seed of tobaccocultivar NCTG-61 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-120314,where the leaf has a reduced amount of nornicotine and/orN′-nitrosonornicotine (NNN).

In an aspect, the present disclosure includes a harvested leaf, or partthereof, produced by growing the seed of tobacco cultivar NCTG-61 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-120314, where the leaf has areduced amount of nornicotine and/or N′-nitrosonornicotine (NNN), wherethe reduced amount of nornicotine and/or N′-nitrosonornicotine (NNN) isreduced in a smoke stream produced from the leaf.

In a further aspect, the present disclosure includes a tobacco product,prepared from a tobacco plant, or part thereof, produced by growing theseed of tobacco cultivar NCTG-61 SRC, a representative sample seed ofthe cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-120314, where the product is selected from the group consistingof pipe tobacco, cigar tobacco, cigarette tobacco, chewing tobacco, leaftobacco, shredded tobacco, and cut tobacco.

In an aspect, the present disclosure includes a tobacco product,prepared from a tobacco plant, or part thereof, produced by growing theseed of tobacco cultivar NCTG-61 SRC, a representative sample seed ofthe cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-120314, where the product is selected from the group consistingof a cigarillo, a non-ventilated recess filter cigarette, a ventedrecess filter cigarette, a cigar, snuff, and chewing tobacco.

In another aspect, the present disclosure includes a tobacco product,prepared from a tobacco plant, or part thereof, produced by growing theseed of tobacco cultivar NCTG-61 SRC, a representative sample seed ofthe cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-120314, where the product is selected from the group consistingof a cigarillo, a non-ventilated recess filter cigarette, a ventedrecess filter cigarette, a cigar, snuff, and chewing tobacco, and wherethe product has a reduced amount of nornicotine and/orN′-nitrosonornicotine (NNN).

In an aspect, the present disclosure includes a part of a tobacco plant,produced by growing the seed of tobacco cultivar NCTG-61 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-120314, where the part is selectedfrom the group consisting of leaf, pollen, ovule, embryo, cotyledon,hypocotyl, meristematic cell, protoplast, root, root tip, pistil,anther, flower, shoot, stem, pod, and petiole.

In another aspect, the present disclosure includes a tissue cultureproduced from a protoplast or cell from a tobacco plant, or partthereof, produced by growing the seed of tobacco cultivar NCTG-61 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-120314, where the cell orprotoplast of the tissue culture is produced from a plant part selectedfrom the group consisting of a leaf, pollen, embryo, cotyledon,hypocotyl, meristematic cell, root, root tip, pistil, anther, flower,shoot, stem, pod, and petiole.

In an aspect, the present disclosure includes a tobacco plantregenerated from a tissue culture produced from a protoplast or cellfrom a tobacco plant, or part thereof, produced by growing the seed oftobacco cultivar NCTG-61 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-120314, where the cell or protoplast of the tissue culture isproduced from a plant part selected from the group consisting of a leaf,pollen, embryo, cotyledon, hypocotyl, meristematic cell, root, root tip,pistil, anther, flower, shoot, stem, pod, and petiole where theregenerated plant has all, or essentially all of the morphological andphysiological characteristics of cultivar NCTG-61 SRC.

In an aspect, the present disclosure includes an F₁ progeny plant oftobacco cultivar NCTG-61 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-120314.

In another aspect, the present disclosure includes an F₁ progeny plantof tobacco cultivar NCTG-61 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-120314, where the F₁ plant is cytoplasmic male sterile (CMS).

In another aspect, the present disclosure includes an F₁ progeny seedproduced by a method of comprising crossing two tobacco plants andharvesting the resultant tobacco seed, where at least one tobacco plantis a tobacco plant produced by growing the seed of tobacco cultivarNCTG-61 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No. PTA-120314, where atleast one tobacco plant is cytoplasmic male sterile (CMS).

In another aspect, the present disclosure includes a method forproducing a tobacco seed comprising crossing two tobacco plants andharvesting the resultant tobacco seed, where at least one tobacco plantis a tobacco plant produced by growing the seed of tobacco cultivarNCTG-61 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No. PTA-120314.

In another aspect, the present disclosure includes a method forproducing a tobacco seed comprising crossing two tobacco plants andharvesting the resultant tobacco seed, where at least one tobacco plantis a tobacco plant produced by growing the seed of tobacco cultivarNCTG-61 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No. PTA-120314, where theplant of tobacco cultivar NCTG-61 SRC is the female parent.

In another aspect, the present disclosure includes a method forproducing a tobacco seed comprising crossing two tobacco plants andharvesting the resultant tobacco seed, where at least one tobacco plantis a tobacco plant produced by growing the seed of tobacco cultivarNCTG-61 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No. PTA-120314, where theplant of tobacco cultivar NCTG-61 SRC is the male parent.

In another aspect, the present disclosure includes a method forproducing a tobacco seed comprising crossing two tobacco plants andharvesting the resultant tobacco seed, where at least one tobacco plantis a tobacco plant produced by growing the seed of tobacco cultivarNCTG-61 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No, PTA-120314, where atleast one tobacco plant is cytoplasmic male sterile (CMS).

In an aspect, the present disclosure also includes a container of F₁progeny seeds produced by a method of comprising crossing two tobaccoplants and harvesting the resultant tobacco seed, where at least onetobacco plant is a tobacco plant produced by growing the seed of tobaccocultivar NCTG-61 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-120314,where at least one tobacco plant is cytoplasmic male sterile (CMS).

In an aspect, the present disclosure includes an F₁ progeny plantproduced by growing a seed produced by a method of comprising crossingtwo tobacco plants and harvesting the resultant tobacco seed, where atleast one tobacco plant is a tobacco plant produced by growing the seedof tobacco cultivar NCTG-61 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-120314, and where at least one tobacco plant is cytoplasmic malesterile (CMS).

In an aspect, the present disclosure also includes a harvested leaf ofan F₁ progeny plant produced by growing a seed produced by a method ofcomprising crossing two tobacco plants and harvesting the resultanttobacco seed, where at least one tobacco plant is a tobacco plantproduced by growing the seed of tobacco cultivar NCTG-61 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-120314, where at least one tobaccoplant is cytoplasmic male sterile (CMS).

In another aspect, the present disclosure further includes a harvestedleaf of an F₁ progeny plant having a reduced amount of nornicotineand/or N′-nitrosonomicotine (NNN) produced by growing a seed produced bya method of comprising crossing two tobacco plants and harvesting theresultant tobacco seed, where at least one tobacco plant is a tobaccoplant produced by growing the seed of tobacco cultivar NCTG-61 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-120314, where at least one tobaccoplant is cytoplasmic male sterile (CMS).

In an aspect, the present disclosure also includes a harvested leaf ofan F₁ progeny plant having a reduced amount of nornicotine and/orN′-nitrosonomicotine (NNN) in a smoke stream produced from the leaf,where the plant is produced by growing a seed produced by a method ofcomprising crossing two tobacco plants and harvesting the resultanttobacco seed, where at least one tobacco plant is a tobacco plantproduced by growing the seed of tobacco cultivar NCTG-61 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-120314, where at least one tobaccoplant is cytoplasmic male sterile (CMS).

In an aspect, the present disclosure includes a tobacco product preparedfrom an F₁ progeny tobacco plant, or part thereof, where the plant orpart thereof is produced by growing a F₁ progeny seed produced by amethod comprising crossing two tobacco plants and harvesting theresultant tobacco seed, where at least one tobacco plant is produced bygrowing a seed of tobacco cultivar NCTG-61 SRC, a representative sampleseed of the cultivar having been deposited with the ATCC under ATCCAccession No. PTA-120314, where at least one tobacco plant iscytoplasmic male sterile, and where the tobacco product is selected fromthe group consisting of pipe tobacco, cigar tobacco, cigarette tobacco,chewing tobacco, leaf tobacco, shredded tobacco, and cut tobacco.

In an aspect, the present disclosure further includes a tobacco productprepared from an F₁ progeny tobacco plant, or part thereof, where theplant or part thereof is produced by growing a F₁ progeny seed producedby a method comprising crossing two tobacco plants and harvesting theresultant tobacco seed, where at least one tobacco plant is produced bygrowing a seed of tobacco cultivar NCTG-61 SRC, a representative sampleseed of the cultivar having been deposited with the ATCC under ATCCAccession No. PTA-120314, where at least one tobacco plant iscytoplasmic male sterile, and where the tobacco product is selected fromthe group consisting of a cigarillo, a non-ventilated recess filtercigarette, a vented recess filter cigarette, a cigar, snuff, and chewingtobacco.

In an aspect, the present disclosure further includes a tobacco productprepared from an F₁ progeny tobacco plant, or part thereof, where theplant or part thereof is produced by growing a F₁ progeny seed producedby a method comprising crossing two tobacco plants and harvesting theresultant tobacco seed, where at least one tobacco plant is produced bygrowing a seed of tobacco cultivar NCTG-61 SRC, a representative sampleseed of the cultivar having been deposited with the ATCC under ATCCAccession No, PTA-120314, where at least one tobacco plant iscytoplasmic male sterile, and where the tobacco product is selected fromthe group consisting of a cigarillo, a non-ventilated recess filtercigarette, a vented recess filter cigarette, a cigar, snuff, and chewingtobacco, and further where the product has a reduced amount ofnornicotine and/or N′-nitrosonornicotine (NNN).

In another aspect, the present disclosure includes a seed of tobaccocultivar CMS NCTG-61 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-13456.

In an aspect, the present disclosure includes a tobacco plant, or partthereof, produced by growing the seed of tobacco cultivar CMS NCTG-61SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13456.

In another aspect, the present disclosure includes a harvested leaf, orpart thereof, of a tobacco plant, or part thereof, produced by growingthe seed of tobacco cultivar CMS NCTG-61 SRC, a representative sampleseed of the cultivar having been deposited with the ATCC under ATCCAccession No. PTA-13456.

In an aspect, the present disclosure includes a harvested leaf, or partthereof, of a tobacco plant, produced by growing the seed of tobaccocultivar CMS NCTG-61 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-13456,where the leaf has a reduced amount of nornicotine and/orN′-nitrosonornicotine (NNN).

In another aspect, the present disclosure includes a harvested leaf, orpart thereof, produced by growing the seed of tobacco cultivar CMSNCTG-61 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No. PTA-13456, where theleaf has a reduced amount of nornicotine and/or N′-nitrosonornicotine(NNN), where the reduced amount of nornicotine and/orN′-nitrosonornicotine (NNN) is reduced in a smoke stream produced fromthe leaf.

In a further aspect, the present disclosure includes a tobacco product,prepared from a tobacco plant, or part thereof, produced by growing theseed of tobacco cultivar CMS NCTG-61 SRC, a representative sample seedof the cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-13456, where the product is selected from the group consistingof pipe tobacco, cigar tobacco, cigarette tobacco, chewing tobacco, leaftobacco, shredded tobacco, and cut tobacco.

In an aspect, the present disclosure includes a tobacco product,prepared from a tobacco plant, or part thereof, produced by growing theseed of tobacco cultivar CMS NCTG-61 SRC, a representative sample seedof the cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-13456, where the product is selected from the group consistingof a cigarillo, a non-ventilated recess filter cigarette, a ventedrecess filter cigarette, a cigar, snuff, and chewing tobacco.

In another aspect, the present disclosure includes a tobacco product,prepared from a tobacco plant, or part thereof, produced by growing theseed of tobacco cultivar CMS NCTG-61 SRC, a representative sample seedof the cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-13456, where the product is selected from the group consistingof a cigarillo, a non-ventilated recess filter cigarette, a ventedrecess filter cigarette, a cigar, snuff, and chewing tobacco where theproduct has a reduced amount of nornicotine and/or N′-nitrosonornicotine(NNN).

In an aspect, the present disclosure includes a part of a tobacco plant,produced by growing the seed of tobacco cultivar CMS NCTG-61 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13456, where the part is selectedfrom the group consisting of leaf, pollen, ovule, embryo, cotyledon,hypocotyl, meristematic cell, protoplast, root, root tip, pistil,anther, flower, shoot, stem, pod, and petiole.

In another aspect, the present disclosure includes a tissue cultureproduced from a protoplast or cell from a tobacco plant, or partthereof, produced by growing the seed of tobacco cultivar CMS NCTG-61SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13456, where the cell orprotoplast of the tissue culture is produced from a plant part selectedfrom the group consisting of a leaf, pollen, embryo, cotyledon,hypocotyl, meristematic cell, root, root tip, pistil, anther, flower,shoot, stem, pod, and petiole.

In an aspect, the present disclosure includes a tobacco plantregenerated from a tissue culture produced from a protoplast or cellfrom a tobacco plant, or part thereof, produced by growing the seed oftobacco cultivar CMS NCTG-61 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13456, where the cell or protoplast of the tissue culture isproduced from a plant part selected from the group consisting of a leaf,pollen, embryo, cotyledon, hypocotyl, meristematic cell, root, root tip,pistil, anther, flower, shoot, stem, pod, and petiole where theregenerated plant has all, or essentially all of the morphological andphysiological characteristics of cultivar CMS NCTG-61 SRC.

In an aspect, the present disclosure includes an F₁ progeny plant oftobacco cultivar CMS NCTG-61 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13456.

In another aspect, the present disclosure includes an F₁ progeny plantof tobacco cultivar CMS NCTG-61 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13456, where the F₁ plant is cytoplasmic male sterile (CMS).

In another aspect, the present disclosure includes an F₁ progeny seedproduced by a method of comprising crossing two tobacco plants andharvesting the resultant tobacco seed, where at least one tobacco plantis cytoplasmic male sterile (CMS), where the cytoplasmic male sterileplant is a plant of tobacco cultivar CMS NCTG-61 SRC, a representativesample seed of the cultivar having been deposited with the ATCC underATCC Accession No. PTA-13456.

In another aspect, the present disclosure includes a method forproducing a tobacco seed comprising crossing two tobacco plants andharvesting the resultant tobacco seed, where at least one tobacco plantis a tobacco plant produced by growing the seed of tobacco cultivar CMSNCTG-61 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No. PTA-13456.

In another aspect, the present disclosure includes a method forproducing a tobacco seed comprising crossing two tobacco plants andharvesting the resultant tobacco seed, where at least one tobacco plantis a tobacco plant produced by growing the seed of tobacco cultivar CMSNCTG-61 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No. PTA-13456, where theplant of tobacco cultivar CMS NCTG-61 SRC is the female parent.

In an aspect, the present disclosure includes a method for producing atobacco seed comprising crossing two tobacco plants and harvesting theresultant tobacco seed, where at least one tobacco plant is cytoplasmicmale sterile, where the cytoplasmic male sterile plant is a plant oftobacco cultivar CMS NCTG-61 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13456.

In an aspect, the present disclosure also includes a container of F₁progeny seeds produced by a method of comprising crossing two tobaccoplants and harvesting the resultant tobacco seed, where at least onetobacco plant is cytoplasmic male sterile (CMS), where the cytoplasmicmale sterile plant is a tobacco plant produced by growing the seed oftobacco cultivar CMS NCTG-61 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13456.

In an aspect, the present disclosure includes an F₁ progeny plantproduced by growing a seed produced by a method of comprising crossingtwo tobacco plants and harvesting the resultant tobacco seed, where atleast one tobacco plant is cytoplasmic male sterile (CMS), where thecytoplasmic male sterile plant is a tobacco plant produced by growingthe seed of tobacco cultivar CMS NCTG-61 SRC, a representative sampleseed of the cultivar having been deposited with the ATCC under ATCCAccession No. PTA-13456.

In an aspect, the present disclosure also includes a harvested leaf ofan F₁ progeny plant produced by growing a seed produced by a method ofcomprising crossing two tobacco plants and harvesting the resultanttobacco seed, where at least one tobacco plant is cytoplasmic malesterile (CMS), where the cytoplasmic male sterile plant is a tobaccoplant produced by growing the seed of tobacco cultivar CMS NCTG-61 SRC,a representative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13456.

In another aspect, the present disclosure further includes a harvestedleaf of an F₁ progeny plant having a reduced amount of nornicotineand/or N′-nitrosonomicotine (NNN) produced by growing a seed produced bya method of comprising crossing two tobacco plants and harvesting theresultant tobacco seed, where at least one tobacco plant is cytoplasmicmale sterile (CMS), where the cytoplasmic male sterile plant is atobacco plant produced by growing the seed of tobacco cultivar CMSNCTG-61 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No. PTA-13456.

In an aspect, the present disclosure also includes a harvested leaf ofan F₁ progeny plant having a reduced amount of nornicotine and/orN′-nitrosonornicotine (NNN) in a smoke stream produced from the leaf,where the plant is produced by growing a seed produced by a method ofcomprising crossing two tobacco plants and harvesting the resultanttobacco seed, where at least one tobacco plant is cytoplasmic malesterile (CMS), where the cytoplasmic male sterile plant is a tobaccoplant produced by growing the seed of tobacco cultivar CMS NCTG-61 SRC,a representative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13456.

In an aspect, the present disclosure includes a tobacco product preparedfrom an F₁ progeny tobacco plant, or part thereof, where the plant orpart thereof is produced by growing a F₁ progeny seed produced by amethod comprising crossing two tobacco plants and harvesting theresultant tobacco seed, where at least one tobacco plant is cytoplasmicmale sterile (CMS), where the cytoplasmic male sterile plant is atobacco plant produced by growing the seed of tobacco cultivar CMSNCTG-61 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No. PTA-13456, and wherethe tobacco product is selected from the group consisting of pipetobacco, cigar tobacco, cigarette tobacco, chewing tobacco, leaftobacco, shredded tobacco, and cut tobacco.

In an aspect, the present disclosure further includes a tobacco productprepared from an F₁ progeny tobacco plant, or part thereof, where theplant or part thereof is produced by growing a F₁ progeny seed producedby a method comprising crossing two tobacco plants and harvesting theresultant tobacco seed, where at least one tobacco plant is cytoplasmicmale sterile (CMS), where the cytoplasmic male sterile plant is atobacco plant produced by growing the seed of tobacco cultivar CMSNCTG-61 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No. PTA-13456, and wherethe tobacco product is selected from the group consisting of acigarillo, a non-ventilated recess filter cigarette, a vented recessfilter cigarette, a cigar, snuff, and chewing tobacco.

In an aspect, the present disclosure further includes a tobacco productprepared from an F₁ progeny tobacco plant, or part thereof, where theplant or part thereof is produced by growing a F₁ progeny seed producedby a method comprising crossing two tobacco plants and harvesting theresultant tobacco seed, where at least one tobacco plant is cytoplasmicmale sterile (CMS), where the cytoplasmic male sterile plant is atobacco plant produced by growing the seed of tobacco cultivar CMSNCTG-61 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No. PTA-13456, and wherethe tobacco product is selected from the group consisting of acigarillo, a non-ventilated recess filter cigarette, a vented recessfilter cigarette, a cigar, snuff, and chewing tobacco, and further wherethe product has a reduced amount of nornicotine and/orN′-nitrosonornicotine (NNN).

In another aspect, the present disclosure includes a seed of hybridtobacco cultivar NC196 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-13458.

In an aspect, the present disclosure includes a tobacco plant, or partthereof, produced by growing a seed of hybrid tobacco cultivar NC196SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13458.

In a further aspect, the present disclosure includes a harvested leaf,or part thereof, of a tobacco plant, or part thereof, produced bygrowing the seed of hybrid tobacco cultivar NC196 SRC, a representativesample seed of the cultivar having been deposited with the ATCC underATCC Accession No. PTA-13458.

In another aspect, the present disclosure includes a harvested leaf, orpart thereof, of a tobacco plant, produced by growing the seed of hybridtobacco cultivar NC196 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-13458,where the leaf has a reduced amount of nornicotine and/orN′-nitrosonornicotine (NNN).

In an aspect, the present disclosure includes a harvested leaf, or partthereof, produced by growing the seed of hybrid tobacco cultivar NC196SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13458, where the leaf has areduced amount of nornicotine and/or N′-nitrosonornicotine (NNN), wherethe reduced amount of nornicotine and/or N′-nitrosonornicotine (NNN) isreduced in a smoke stream produced from the leaf.

In a further aspect, the present disclosure includes a tobacco product,prepared from a tobacco plant, or part thereof, produced by growing theseed of hybrid tobacco cultivar NC196 SRC, a representative sample seedof the cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-13458, where the product is selected from the group consistingof pipe tobacco, cigar tobacco, cigarette tobacco, chewing tobacco, leaftobacco, shredded tobacco, and cut tobacco.

In an aspect, the present disclosure includes a tobacco product,prepared from a tobacco plant, or part thereof, produced by growing theseed of hybrid tobacco cultivar NC196 SRC, a representative sample seedof the cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-13458, where the product is selected from the group consistingof a cigarillo, a non-ventilated recess filter cigarette, a ventedrecess filter cigarette, a cigar, snuff, and chewing tobacco.

In another aspect, the present disclosure includes a tobacco product,prepared from a tobacco plant, or part thereof, produced by growing theseed of hybrid tobacco cultivar NC196 SRC, a representative sample seedof the cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-13458, where the product is selected from the group consistingof a cigarillo, a non-ventilated recess filter cigarette, a ventedrecess filter cigarette, a cigar, snuff, and chewing tobacco where theproduct has a reduced amount of nornicotine and/or N′-nitrosonornicotine(NNN).

In an aspect, the present disclosure includes a part of a tobacco plant,produced by growing a seed of hybrid tobacco cultivar NC196 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13458, where the part is selectedfrom the group consisting of leaf, pollen, ovule, embryo, cotyledon,hypocotyl, meristematic cell, protoplast, root, root tip, pistil,anther, flower, shoot, stem, pod, and petiole.

In a further aspect, the present disclosure includes a tissue cultureproduced from a protoplast or cell from a tobacco plant, or partthereof, produced by growing the seed of hybrid tobacco cultivar NC196SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No, PTA-13458, where the cell orprotoplast of the tissue culture is produced from a plant part selectedfrom the group consisting of a leaf, pollen, embryo, cotyledon,hypocotyl, meristematic cell, root, root tip, pistil, anther, flower,shoot, stem, pod, and petiole.

In an aspect, the present disclosure includes a tobacco plantregenerated from a tissue culture produced from a protoplast or cellfrom a tobacco plant, or part thereof, produced by growing the seed ofhybrid tobacco cultivar NC196 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13458, where the cell or protoplast of the tissue culture isproduced from a plant part selected from the group consisting of a leaf,pollen, embryo, cotyledon, hypocotyl, meristematic cell, root, root tip,pistil, anther, flower, shoot, stem, pod, and petiole, where theregenerated plant has all, or essentially all of the morphological andphysiological characteristics of hybrid cultivar NC196 SRC.

In an aspect, the present disclosure includes an F₁ progeny plant ofhybrid tobacco cultivar NC196 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13458.

In an aspect, the present disclosure includes an F₁ progeny plant ofhybrid tobacco cultivar NC196 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13458, where the plant of tobacco cultivare NC196 SRC is the femaleparent.

In an aspect, the present disclosure includes an F₁ progeny plant ofhybrid tobacco cultivar NC196 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13458, where the plant of tobacco cultivare NC196 SRC is the maleparent.

In another aspect, the present disclosure includes an F₁ progeny plantof hybrid tobacco cultivar NC196 SRC, a representative sample seed ofthe hybrid cultivar having been deposited with the ATCC under ATCCAccession No. PTA-13458, where the F₁ plant is cytoplasmic male sterile(CMS).

In another aspect, the present disclosure includes a method forproducing a tobacco seed of NC196 SRC comprising crossing two tobaccoplants and harvesting the resultant tobacco seed, where one tobaccoplant is a tobacco plant produced by growing the seed of tobaccocultivar NC 1562-1 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-13457.

In another aspect, the present disclosure includes a method forproducing a tobacco seed of NC196 SRC comprising crossing two tobaccoplants and harvesting the resultant tobacco seed, where one tobaccoplant is a tobacco plant produced by growing the seed of tobaccocultivar CMS NCTG-61 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-13456.

In another aspect, the present disclosure includes a method forproducing a tobacco seed of NC196 SRC comprising crossing two tobaccoplants and harvesting the resultant tobacco seed, where one tobaccoplant is a tobacco plant produced by growing the seed of tobaccocultivar CMS NCTG-61 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No, PTA-13456and a second tobacco plant is a tobacco plant produced by growing theseed of tobacco cultivar NC1562-1 SRC, a representative sample seed ofthe cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-13457.

In an aspect, the present disclosure includes a method of vegetativelypropagating a plant of a tobacco cultivar comprising the steps of (a)collecting tissue capable of being propagated from a plant of a tobaccocultivar selected from the group consisting of K326 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13455, CMS K326 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13454, K346 SRC, a representativesample seed of the cultivar having been deposited with the ATCC underATCC Accession No. PTA-13453, CMS K346 SRC, a representative sample seedof the cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-13452, NC1562-1 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13457, NCTG-61 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-120314,CMS NCTG-61 SRC, a representative sample seed of the cultivar havingbeen deposited with the ATCC under ATCC Accession No. PTA-13456, andhybrid cultivar NC196 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-13458;(b) cultivating the tissue to obtain a proliferated shoot; and (c)rooting the proliferated shoots to obtain a rooted plantlet.

In an aspect, the present disclosure includes a method of vegetativelypropagating a plant of a tobacco cultivar comprising the steps of (a)collecting tissue capable of being propagated from a plant of a tobaccocultivar selected from the group consisting of K326 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13455, CMS K326 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13454, K346 SRC, a representativesample seed of the cultivar having been deposited with the ATCC underATCC Accession No. PTA-13453, CMS K346 SRC, a representative sample seedof the cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-13452, NC1562-1 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13457, NCTG-61 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-120314,CMS NCTG-61 SRC, a representative sample seed of the cultivar havingbeen deposited with the ATCC under ATCC Accession No. PTA-13456, andhybrid cultivar NC196 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-13458;(b) cultivating the tissue to obtain a proliferated shoot; (c) rootingthe proliferated shoots to obtain a rooted plantlet; and (d) growing aplant from the rooted plantlet.

In an aspect, the present disclosure includes a method of introducing adesired trait into a tobacco cultivar comprising: (a) crossing a plantof a first tobacco cultivar selected from the group consisting of K326SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13455, CMS K326 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13454, K346 SRC, a representativesample seed of the cultivar having been deposited with the ATCC underATCC Accession No. PTA-13453, CMS K346 SRC, a representative sample seedof the cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-13452, NC1562-1 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13457, NCTG-61 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-120314,and CMS NCTG-61 SRC, a representative sample seed of the cultivar havingbeen deposited with the ATCC under ATCC Accession No. PTA-13456, with asecond tobacco plant that comprises a desired trait to produce an F₁progeny seed; (b) growing the F₁ progeny seed and selecting an F₁progeny plant that comprises the desired trait; (c) crossing theselected F₁ progeny plant with a plant of said first tobacco cultivarselected from the group consisting of K326 SRC, a representative sampleseed of the cultivar having been deposited with the ATCC under ATCCAccession No, PTA-13455, CMS K326 SRC, a representative sample seed ofthe cultivar having been deposited with the ATCC under ATCC AccessionNo, PTA-13454, K346 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No, PTA-13453,CMS K346 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No. PTA-13452, NC1562-1SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13457, NCTG-61 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-120314, and CMS NCTG-61 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13456 to produce a backcross BC₁F₁progeny seed; (d) growing the BC₁F₁ progeny seed and selecting abackcross BC₁F₁ progeny plant comprising the desired trait andessentially all of the physiological and morphological characteristicsof said first tobacco cultivar, K326 SRC, CMS K326 SRC, K346 SRC, CMSK346 SRC, NC1562-1 SRC, NCTG-61 SRC, or CMS NCTG-61 SRC; and (e)repeating steps (c) and (d) three or more times in succession to produceselected fourth or higher backcross progeny that comprise the desiredtrait.

In an aspect, the present disclosure includes a method of introducing adesired trait into a tobacco cultivar comprising: (a) crossing a plantof a first tobacco cultivar selected from the group consisting of K326SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13455, CMS K326 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13454, K346 SRC, a representativesample seed of the cultivar having been deposited with the ATCC underATCC Accession No, PTA-13453, CMS K346 SRC, a representative sample seedof the cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-13452, NC1562-1 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13457, NCTG-61 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-120314,and CMS NCTG-61 SRC, a representative sample seed of the cultivar havingbeen deposited with the ATCC under ATCC Accession No. PTA-13456, with asecond tobacco plant that comprises a desired trait to produce an F₁progeny seed; (b) growing the F₁ progeny seed and selecting an F₁progeny plant that comprises the desired trait; (c) crossing theselected F₁ progeny plant with a plant of the first tobacco cultivarselected from the group consisting of K326 SRC, a representative sampleseed of the cultivar having been deposited with the ATCC under ATCCAccession No. PTA-13455, K346 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13453, and NC1562-1 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13457, NCTG-61 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-120314,and CMS NCTG-61 SRC, a representative sample seed of the cultivar havingbeen deposited with the ATCC under ATCC Accession No. PTA-13456 toproduce a backcross BC₁F₁ progeny seed; (d) growing the BC₁F₁F₂ progenyseed and selecting a backcross BC₁F₁ progeny plant comprising thedesired trait and essentially all of the physiological and morphologicalcharacteristics of the first tobacco cultivar, K326 SRC, CMS K326 SRC,K346 SRC, CMS K346 SRC, NC1562-1 SRC, NCTG-61 SRC, or CMS NCTG-61 SRC;and (e) repeating steps (c) and (d) three or more times in succession toproduce selected fourth or higher backcross progeny that comprise thedesired trait, where the trait is cytoplasmic male sterility (CMS).

In an aspect, the present disclosure includes a method of introducing adesired trait into a tobacco cultivar comprising: (a) crossing a plantof a first tobacco cultivar selected from the group consisting of K326SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13455, K346 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13453, NCTG-61 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-120314, and NC1562-1 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13457, with a second tobacco plantthat comprises a desired trait to produce an F₁ progeny seed; (b)growing the F₁ progeny seed and selecting an F₁ progeny plant thatcomprises the desired trait; (c) crossing the selected F₁ progeny plantwith a plant of the first tobacco cultivar selected from the groupconsisting of K326 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-13455,K346 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No. PTA-13453, NCTG-61 SRC,a representative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-120314, and NC1562-1 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13457 to produce a backcross BC₁F₁progeny seed; (d) growing the BC₁F₁ progeny seed and selecting abackcross BC₁F₁ progeny plant comprising the desired trait and thephysiological and essentially all of morphological characteristics ofthe first tobacco cultivar K326 SRC, K346 SRC, NCTG-61 SRC, or NC1562-1SRC; and (e) repeating steps (c) and (d) three or more times insuccession to produce selected fourth or higher backcross progeny thatcomprise the desired trait, where the trait is cytoplasmic malesterility (CMS) and the CMS trait is obtained from the cytoplasm ofNicotiana suaveolens or Nicotiana glauca.

In an aspect, the present disclosure includes a method of introducing adesired trait into a tobacco cultivar comprising: (a) crossing a plantof a first tobacco cultivar selected from the group consisting of K326SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No, PTA-13455, K346 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13453, and NC1562-1 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13457, and NCTG-61 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No, PTA-120314, with a second tobaccoplant that comprises a desired trait to produce an F₁ progeny seed; (b)growing the F₁ progeny seed and selecting an F₁ progeny plant thatcomprises the desired trait; (c) crossing the selected F₁ progeny plantwith a plant of the first tobacco cultivar selected from the groupconsisting of K326 SRC, K346 SRC, NCTG-61 SRC, and NC1562-1 SRC toproduce a backcross BC₁F₁ progeny seed; (d) growing the BC₁F₁ progenyseed and selecting a backcross BC₁F₁ progeny plant comprising thedesired trait and essentially all of the physiological and morphologicalcharacteristics of tobacco cultivar K326 SRC, K346 SRC, NCTG-61 SRC, orNC1562-1 SRC; and (e) repeating steps (c) and (d) three or more times insuccession to produce selected fourth or higher backcross progeny thatcomprise the desired trait, where the trait is cytoplasmic malesterility (CMS) and the CMS trait is obtained from the cytoplasm ofNicotiana suaveolens, and where the second tobacco plant is selectedfrom the group consisting of CMS K326 SRC, CMS K346 SRC, and CMS NCTG-61SRC.

In an aspect, the present disclosure includes a tobacco plant producedby a method comprising introducing a desired trait into a tobaccocultivar comprising: (a) crossing a plant of a first tobacco cultivarselected from the group consisting of K326 SRC, a representative sampleseed of the cultivar having been deposited with the ATCC under ATCCAccession No. PTA-13455, CMS K326 SRC, a representative sample seed ofthe cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-13454, K346 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-13453,CMS K346 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No. PTA-13452, NC1562-1SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13457, NCTG-61 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-120314, and CMS NCTG-61 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13456, with a second tobacco plantthat comprises a desired trait to produce an F₁ progeny seed; (b)growing the F₁ progeny seed and selecting an F₁ progeny plant thatcomprises the desired trait; (c) crossing the selected F₁ progeny plantwith a plant of the first tobacco cultivar selected from the groupconsisting of K326 SRC, CMS K326 SRC, K346 SRC, CMS K346 SRC, NC1562-1SRC, NCTG-61 SRC, and CMS NCTG-61 SRC to produce a backcross BC₁F₁progeny seed; (d) growing the BC₁F₁ progeny seed and selecting abackcross BC₁F₁ progeny plant comprising the desired trait andessentially all of the physiological and morphological characteristicsof the first tobacco cultivar K326 SRC, CMS K326 SRC, K346 SRC, CMS K346SRC, NC1562-1 SRC, NCTG-61 SRC, or CMS NCTG-61 SRC; and (e) repeatingsteps (c) and (d) three or more times in succession to produce selectedfourth or higher backcross progeny that comprise the desired trait.

In another aspect, the present disclosure includes a method ofintroducing a desired trait into a tobacco cultivar comprising: (a)crossing a plant of a first tobacco cultivar selected from the groupconsisting of K326 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-13455,CMS K326 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No. PTA-13454, K346 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13453, CMS K346 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13452, NC1562-1 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No, PTA-13457, NCTG-61 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-120314, and CMS NCTG-61 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13456, with a plant of a secondtobacco cultivar that comprises a desired trait to produce a progenyplant where the desired trait is selected from the group consisting ofherbicide resistance, pest resistance, disease resistance, high yield,high grade index, curability, curing quality, mechanical harvestability,holding ability, leaf quality, height, plant maturation, early maturing,early to medium maturing, medium maturing, medium to late maturing, latematuring; small stalk, medium stalk, large stalk, leaf number per plant,5-10 leaves per plant, 11-15 leaves per plant, 16-21 leaves per plant,and any combination thereof, to produce an F₁ progeny seed; (b) growingthe F₁ progeny seed into an F₁ progeny plant and selecting the F₁progeny plant having the desired trait; (c) crossing the selected F₁progeny plant with a plant of the first tobacco cultivar selected fromthe group consisting of K326 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13455, CMS K326 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-13454,K346 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No. PTA-13453, CMS K346SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No, PTA-13452, NC1562-1 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13457, NCTG-61 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-120314, and CMS NCTG-61 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13456 to produce a backcrossprogeny plant seed; (d) growing the backcross progeny plant seed into abackcross progeny plant and selecting the backcross progeny plantcomprising the desired trait and essentially all of the physiologicaland morphological characteristics of the first tobacco cultivar, K326SRC, CMS K326 SRC, K346 SRC, CMS K346 SRC, NC1562-1 SRC, NCTG-61 SRC, orCMS NCTG-61 SRC; and (e) repeating steps (c) and (d) one or more timesin succession to produce a selected fourth or higher backcross progenyplant that comprises the desired trait and essentially all of thephysiological and morphological characteristics of the first tobaccocultivar, K326 SRC, CMS K326 SRC, K346 SRC, CMS K346 SRC, NC1562-1 SRC,NCTG-61 SRC, or CMS NCTG-61 SRC.

In another aspect, the present disclosure includes a method ofintroducing a desired trait into a tobacco cultivar comprising: (a)crossing a plant of a first tobacco cultivar selected from the groupconsisting of K326 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-13455,CMS K326 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No. PTA-13454, K346 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13453, CMS K346 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13452, NC1562-1 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13457, NCTG-61 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-120314, and CMS NCTG-61 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13456, with a plant of a secondtobacco cultivar that comprises a desired trait to produce a progenyplant where the desired trait is selected from the group consisting ofherbicide resistance, pest resistance, disease resistance, high yield,high grade index, curability, curing quality, mechanical harvestability,holding ability, leaf quality, height, plant maturation, early maturing,early to medium maturing, medium maturing, medium to late maturing, latematuring; small stalk, medium stalk, large stalk, leaf number per plant,5-10 leaves per plant, 11-15 leaves per plant, 16-21 leaves per plant,and any combination thereof, to produce an F₁ progeny seed; (b) growingthe F₁ progeny seed into an F₁ progeny plant and selecting the F₁progeny plant having the desired trait; (c) crossing the selected F₁progeny plant with a plant of the first tobacco cultivar selected fromthe group consisting of K326 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13455, CMS K326 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-13454,K346 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No. PTA-13453, CMS K346SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13452, NC1562-1 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13457, NCTG-61 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-120314, and CMS NCTG-61 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13456 to produce a backcrossprogeny plant seed; (d) growing the backcross progeny plant seed into abackcross progeny plant and selecting the backcross progeny plantcomprising the desired trait and essentially all of the physiologicaland morphological characteristics of a tobacco cultivar, K326 SRC, CMSK326 SRC, K346 SRC, CMS K346 SRC, NC1562-1 SRC, NCTG-61 SRC, or CMSNCTG-61 SRC; and (e) repeating steps (c) and (d) one or more times insuccession to produce a selected fourth or higher backcross progenyplant that comprises the desired trait and essentially all of thephysiological and morphological characteristics of the first tobaccocultivar, K326 SRC, CMS K326 SRC, K346 SRC, CMS K346 SRC, NC1562-1 SRC,NCTG-61 SRC, or CMS NCTG-61 SRC, where the plant has a desired trait.

In another aspect, the present disclosure includes a method ofintroducing a desired trait into a tobacco cultivar comprising: (a)crossing a plant of a first tobacco cultivar selected from the groupconsisting of K326 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No, PTA-13455,CMS K326 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No. PTA-13454, K346 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13453, CMS K346 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13452, NC1562-1 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13457, NCTG-61 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-120314, and CMS NCTG-61 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13456, with a plant of a secondtobacco cultivar that comprises a desired trait to produce a progenyplant where the desired trait is selected from the group consisting ofherbicide resistance, pest resistance, disease resistance, high yield,high grade index, curability, curing quality, mechanical harvestability,holding ability, leaf quality, height, plant maturation, early maturing,early to medium maturing, medium maturing, medium to late maturing, latematuring; small stalk, medium stalk, large stalk, leaf number per plant,5-10 leaves per plant, 11-15 leaves per plant, 16-21 leaves per plant,and any combination thereof, to produce an F₁ progeny seed; (b) growingthe F₁ progeny seed into an F₁ progeny plant and selecting the F₁progeny plant having the desired trait; (c) crossing the selected F₁progeny plant with a plant of the first tobacco cultivar selected fromthe group consisting of K326 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13455, CMS K326 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No, PTA-13454,K346 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No, PTA-13453, CMS K346SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13452, NC1562-1 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13457, NCTG-61 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-120314, and CMS NCTG-61 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13456 to produce a backcrossprogeny plant seed; (d) growing the backcross progeny plant seed into abackcross progeny plant and selecting the backcross progeny plantcomprising the desired trait and essentially all of the physiologicaland morphological characteristics of the first tobacco cultivar, K326SRC, CMS K326 SRC, K346 SRC, CMS K346 SRC, NC1562-1 SRC, NCTG-61 SRC, orCMS NCTG-61 SRC; and (e) repeating steps (c) and (d) one or more timesin succession to produce a selected fourth or higher backcross progenyplant that comprises the desired trait and essentially all of thephysiological and morphological characteristics of a tobacco cultivar,K326 SRC, CMS K326 SRC, K346 SRC, CMS K346 SRC, NC1562-1 SRC, NCTG-61SRC, or CMS NCTG-61 SRC, where the plant has a desired trait of diseaseresistance.

In another aspect, the present disclosure includes a method forproducing a tobacco plant having decreased nicotine conversioncomprising: identifying a first tobacco plant having the sequenceselected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ IDNO: 11, and combinations thereof; crossing the first tobacco plant witha second tobacco plant and collecting an F₁ seed; crossing a plant grownfrom the F₁ seed to a third tobacco plant and collecting a secondtobacco seed; and identifying a second tobacco seed or a plant grownfrom the second seed that is homozygous for the sequence selected fromthe group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 11, andcombinations thereof.

In an aspect, the present disclosure includes a method for producing atobacco plant having decreased nicotine conversion comprising:identifying a first tobacco plant having the sequence selected from thegroup consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 11, andcombinations thereof; crossing the first tobacco plant with a secondtobacco plant and collecting an F₁ seed; crossing a plant grown from theF₁ seed to a third tobacco plant and collecting a second tobacco seed;and identifying a second tobacco seed or a plant grown from the secondseed that is homozygous for the sequence selected from the groupconsisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 11, andcombinations thereof, where the second tobacco plant has a sequenceselected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ IDNO: 11, and combinations thereof.

In an aspect, the present disclosure includes a method for producing atobacco plant having decreased nicotine conversion comprising:identifying a first tobacco plant having the sequence selected from thegroup consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 11, andcombinations thereof; crossing the first tobacco plant with a secondtobacco plant and collecting an F₁ seed; crossing a plant grown from theF₁ seed to a third tobacco plant and collecting a second tobacco seed;and identifying a second tobacco seed or a plant grown from the secondseed that is homozygous for a sequence selected from the groupconsisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 11, andcombinations thereof, where the second tobacco plant does not have thesequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO: 11, and combinations thereof, and the third tobacco plantis a tobacco plant having a sequence selected from the group consistingof SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 11, and combinations thereof.

In an aspect, the present disclosure includes a method for producing atobacco plant having decreased nicotine conversion comprising:identifying a first tobacco plant having the sequence selected from thegroup consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 11, andcombinations thereof; crossing the first tobacco plant with a secondtobacco plant and collecting an F₁ seed; crossing a plant grown from theF₁ seed to a third tobacco plant and collecting a second tobacco seed;and identifying a second tobacco seed or a plant grown from the secondseed that is homozygous for a sequence selected from the groupconsisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 11, andcombinations thereof, where the first tobacco plant has the sequence setforth in SEQ ID NO: 1 or SEQ ID NO: 2.

In an aspect, the present disclosure includes a method for producing atobacco plant having decreased nicotine conversion comprising:identifying a first tobacco plant having a sequence selected from thegroup consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 11, andcombinations thereof; crossing the first tobacco plant with a secondtobacco plant and collecting an F₁ seed; crossing a plant grown from theF₁ seed to a third tobacco plant and collecting a second tobacco seed;and identifying a second tobacco seed or a plant grown from the secondseed that is homozygous for a sequence selected from the groupconsisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 11, andcombinations thereof, where the third tobacco plant is a tobacco planthaving a sequence selected from the group consisting of SEQ ID NO: 1,SEQ ID NO: 2, SEQ ID NO: 11, and combinations thereof.

In an aspect, the present disclosure includes a method for producing atobacco plant having decreased nicotine conversion comprising:identifying a first tobacco plant having a sequence selected from thegroup consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 11, andcombinations thereof; crossing the first tobacco plant with a secondtobacco plant and collecting an F₁ seed; crossing a plant grown from theF₁ seed to a third tobacco plant and collecting a second tobacco seed;and identifying a second tobacco seed or a plant grown from the secondseed that is homozygous for a sequence selected from the groupconsisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 11, andcombinations thereof, where the first tobacco plant is a plant of atobacco cultivar selected from the group consisting of K326 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13455, CMS K326 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13454, K346 SRC, a representativesample seed of the cultivar having been deposited with the ATCC underATCC Accession No. PTA-13453, CMS K346 SRC, a representative sample seedof the cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-13452, NC1562-1 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13457, NCTG-61 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-120314,CMS NCTG-61 SRC, a representative sample seed of the cultivar havingbeen deposited with the ATCC under ATCC Accession No. PTA-13456, andhybrid cultivar NC196 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-13458.

In an aspect, the present disclosure includes a method for producing atobacco plant having decreased nicotine conversion comprising:identifying a first tobacco plant having a sequence selected from thegroup consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 11, andcombinations thereof; crossing the first tobacco plant with a secondtobacco plant and collecting an F₁ seed; crossing a plant grown from theF₁ seed to a third tobacco plant and collecting a second tobacco seed;and identifying a second tobacco seed or a plant grown from the secondseed that is homozygous for a sequence selected from the groupconsisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 11, andcombinations thereof, where the third tobacco plant is a plant of atobacco cultivar selected from the group consisting of K326 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13455, CMS K326 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13454, K346 SRC, a representativesample seed of the cultivar having been deposited with the ATCC underATCC Accession No. PTA-13453, CMS K346 SRC, a representative sample seedof the cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-13452, NC1562-1 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13457, NCTG-61 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-120314,CMS NCTG-61 SRC, a representative sample seed of the cultivar havingbeen deposited with the ATCC under ATCC Accession No. PTA-13456, andhybrid cultivar NC196 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No, PTA-13458.

In another aspect, the present disclosure includes a method of producinga plant of a tobacco cultivar selected from the group consisting of K326SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13455, CMS K326 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13454, K346 SRC, a representativesample seed of the cultivar having been deposited with the ATCC underATCC Accession No. PTA-13453, CMS K346 SRC, a representative sample seedof the cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-13452, NC1562-1 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13457, NCTG-61 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-120314,and CMS NCTG-61 SRC, a representative sample seed of the cultivar havingbeen deposited with the ATCC under ATCC Accession No. PTA-13456 havingan additional desired trait comprising the steps of: (a) collectingtissue capable of being propagated from a plant of a tobacco cultivarselected from the group consisting of K326 SRC, a representative sampleseed of the cultivar having been deposited with the ATCC under ATCCAccession No. PTA-13455, CMS K326 SRC, a representative sample seed ofthe cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-13454, K346 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-13453,CMS K346 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No. PTA-13452, NC1562-1SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13457, NCTG-61 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No, PTA-120314, and CMS NCTG-61 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13456; and (b) introducing atransgene conferring the desired trait into the tissue.

In another aspect, the present disclosure includes a method of producingan herbicide resistant tobacco plant comprising transforming a tobaccoplant, or part thereof, produced by growing a seed of a tobacco cultivarselected from the group consisting of K326 SRC, a representative sampleseed of the cultivar having been deposited with the ATCC under ATCCAccession No. PTA-13455, CMS K326 SRC, a representative sample seed ofthe cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-13454, K346 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No, PTA-13453,CMS K346 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No. PTA-13452, NC1562-1SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13457, NCTG-61 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-120314, CMS NCTG-61 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13456, and hybrid cultivar NC196SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13458 with a transgene wherethe transgene confers resistance to an herbicide selected from the groupconsisting of imidazolinone, cyclohexanedione, sulfonylurea, glyphosate,glufosinate, phenoxy proprionic acid, L-phosphinothricin, triazine, andbenzonitrile.

In another aspect, the present disclosure includes an herbicideresistant tobacco plant produced by a method comprising transforming atobacco plant, or part thereof, produced by growing a seed of a tobaccocultivar selected from the group consisting of K326 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13455, CMS K326 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13454, K346 SRC, a representativesample seed of the cultivar having been deposited with the ATCC underATCC Accession No. PTA-13453, CMS K346 SRC, a representative sample seedof the cultivar having been deposited with the ATCC under ATCC AccessionNo, PTA-13452, NC1562-1 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No,PTA-13457, NCTG-61 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-120314,CMS NCTG-61 SRC, a representative sample seed of the cultivar havingbeen deposited with the ATCC under ATCC Accession No. PTA-13456, andhybrid cultivar NC196 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-13458with a transgene where the transgene confers resistance to an herbicideselected from the group consisting of imidazolinone, cyclohexanedione,sulfonylurea, glyphosate, glufosinate, phenoxy proprionic acid,L-phosphinothricin, triazine, and benzonitrile.

In another aspect, the present disclosure includes a method of producinga pest or insect resistant tobacco plant where the method comprisestransforming a tobacco plant produced by growing a seed of a tobaccocultivar selected from the group consisting of K326 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13455, CMS K326 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13454, K346 SRC, a representativesample seed of the cultivar having been deposited with the ATCC underATCC Accession No. PTA-13453, CMS K346 SRC, a representative sample seedof the cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-13452, NC1562-1 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13457, NCTG-61 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-120314,CMS NCTG-61 SRC, a representative sample seed of the cultivar havingbeen deposited with the ATCC under ATCC Accession No. PTA-13456, andhybrid cultivar NC196 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-13458with a transgene that confers pest or insect resistance.

In a further aspect, the present disclosure includes a pest or insectresistant tobacco plant produced by a method comprising transforming atobacco plant produced by growing a seed of a tobacco cultivar selectedfrom the group consisting of K326 SRC, a representative sample seed ofthe cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-13455, CMS K326 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13454, K346 SRC, a representative sample seed of the cultivar havingbeen deposited with the ATCC under ATCC Accession No. PTA-13453, CMSK346 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No. PTA-13452, NC1562-1SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13457, NCTG-61 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-120314, CMS NCTG-61 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13456, and hybrid cultivar NC196SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13458 with a transgene thatconfers pest or insect resistance.

In a further aspect, the present disclosure includes a pest or insectresistant tobacco plant produced by a method comprising transforming atobacco plant produced by growing a seed of a tobacco cultivar selectedfrom the group consisting of K326 SRC, a representative sample seed ofthe cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-13455, CMS K326 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13454, K346 SRC, a representative sample seed of the cultivar havingbeen deposited with the ATCC under ATCC Accession No. PTA-13453, CMSK346 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No. PTA-13452, NC1562-1SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13457, NCTG-61 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-120314, CMS NCTG-61 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13456, and hybrid cultivar NC196SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13458, with a transgene thatconfers pest or insect resistance, where the transgene encodes aBacillus thuringiensis (BT) endotoxin.

In another aspect, the present disclosure includes a method of producinga disease resistant tobacco plant where the method comprisestransforming a tobacco plant produced by growing a seed of a tobaccocultivar selected from the group consisting of K326 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13455, CMS K326 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13454, K346 SRC, a representativesample seed of the cultivar having been deposited with the ATCC underATCC Accession No. PTA-13453, CMS K346 SRC, a representative sample seedof the cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-13452, NC1562-1 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13457, NCTG-61 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-120314,CMS NCTG-61 SRC, a representative sample seed of the cultivar havingbeen deposited with the ATCC under ATCC Accession No. PTA-13456, andhybrid cultivar NC196 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-13458with a transgene that confers disease resistance.

In a further aspect, the present disclosure includes a disease resistanttobacco plant produced by a method comprising transforming a tobaccoplant produced by growing a seed of a tobacco cultivar selected from thegroup consisting of K326 SRC, a representative sample seed of thecultivar having been deposited with the ATCC under ATCC Accession No.PTA-13455, CMS K326 SRC, a representative sample seed of the cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-13454,K346 SRC, a representative sample seed of the cultivar having beendeposited with the ATCC under ATCC Accession No. PTA-13453, CMS K346SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13452, NC1562-1 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No, PTA-13457, NCTG-61 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-120314, CMS NCTG-61 SRC, arepresentative sample seed of the cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13456, and hybrid cultivar NC196SRC, a representative sample seed of the cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13458 with a transgene thatconfers disease resistance.

BRIEF DESCRIPTION OF THE SEQUENCES

SEQ ID NO: 1 sets forth a cyp82e4 W329Stop nucleotide sequence.

SEQ ID NO: 2 sets forth a cyp82e5v2 W422Stop nucleotide sequence.

SEQ ID NO: 3 sets forth a cyp82e4 W329Stop amino acid sequence.

SEQ ID NO: 4 sets forth a cyp82e5v2 W422Stop amino acid sequence.

SEQ ID NO: 5 sets forth a CYP82E4 wild-type nucleotide sequence.

SEQ ID NO: 6 sets forth a CYP82E5v2 wild-type nucleotide sequence.

SEQ ID NO: 7 sets forth a CYP82E4 wild-type amino acid sequence.

SEQ ID NO: 8 sets forth a CYP82E5v2 wild-type amino acid sequence.

SEQ ID NO: 9 sets forth a CYP82E10 wild-type nucleotide sequence.

SEQ ID NO: 10 sets forth a CYP82E10 wild-type amino acid sequence.

SEQ ID NO: 11 sets forth a CYP82E10 P381S nucleotide sequence.

SEQ ID NO: 12 sets forth a CYP82E10 P381S amino acid sequence.

All three tobacco Nicotine Demethylase genes (CYP82E4, CYP82E5v2,CYP82E10) share a common structure: a 939 bp exon 1 and a 612 bp exon 2separated by a large intron, whose length varies among the three genes.See Lewis et al., “Three nicotine demethylase genes mediate nornicotinebiosynthesis in Nicotiana tabacum L.: Functional characterization of theCYP82E10 gene,” Phytochemistry, 71 (2010), 1988-1998. SEQ ID NOs: 1, 2,5, 6, 9, and 11 set forth wild-type or mutant versions of codingsequences of CYP82E4, CYP82E5v2, and CYP82E10. It is understood that,used herein, a plant comprising, having, or homozygous for a sequenceselected from SEQ ID NOs: 1, 2, 5, 6, 9, and 11 refers to a plantcomprising at the CYP82E4, CYP82E5v2, or CYP82E10 endogenous locus agenomic sequence comprising the coding sequence of SEQ ID NO: 1, 2, 5,6, 9, or 11.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1: Comparisons between K326 SRC (shown as “K 326 e4e5e10 Mutant”)and K326 for their NNN concentrations in tobacco filler and cigarettesmoke, and their percent nicotine conversions in tobacco filler. K326SRC has a lower level of NNN compared to K326 in both tobacco filler(˜0.22 versus ˜0.53 ppm, shown as “Filler NNN”) and cigarette smoke(˜0.015 versus ˜0.04 ng/mg Total Particle Matter (TPM), shown as “SmokeNNN”). Percent nicotine conversion is lower in K326 SRC tobacco filler(˜0.8%) than K326 tobacco filler (˜3.1%, shown as “Filler %Conversion”).

DETAILED DESCRIPTION

Unless defined otherwise, technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art. One skilled in the art will recognize many methods can be usedin the practice of the present disclosure. Indeed, the presentdisclosure is in no way limited to the methods and materials described.For purposes of the present disclosure, the following terms are definedbelow.

Any references cited herein are incorporated by reference in theirentireties.

As used herein, the singular form “a,” “an,’ and “the” include pluralreferences unless the context clearly dictates otherwise. For example,the term “a compound” or “at least one compound” may include a pluralityof compounds, including mixtures thereof.

The term “about” is used herein to mean approximately, roughly, around,or in the region of. When the term “about” is used in conjunction with anumerical range, it modifies that range by extending the boundariesabove and below the numerical values set forth.

“Tobacco product” is defined as “any product made or derived fromtobacco that is intended for human use or consumption, including anycomponent, part, or accessory of a tobacco product (except for rawmaterials other than tobacco used in manufacturing a component, part, oraccessory of a tobacco product)” (section 201 of the FD&C Act; 21 U.S.C.321). The label and packaging is part of a tobacco product.

Terms “nicotine conversion rate,” “percent nicotine conversion,” and“percentage nicotine conversion” are used interchangeably. Percentnicotine demethylation in a sample is calculated by dividing the levelof nornicotine by the combined level of nicotine and nornicotine asmeasured in the sample, and multiplying by 100.

K326 SRC

The present disclosure also provides tobacco cultivars, and partsthereof, from K326 SRC, representative sample seeds of this cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-13455.The present disclosure also includes a tobacco plant, or part thereof,produced by growing a seed of K326 SRC. A plant of the presentdisclosure can further include a plant with all, or essentially all ofthe morphological and physiological characteristics of cultivar K326SRC.

While not being limited by process, K326 SRC is a result of theintroduction of three mutated CYP82 genes in a flue-cured tobaccocultivar K326. The three genes are a mutated CYP82E4 gene recited as325-6 #775 in Lewis et al. (“Three nicotine demethylase genes mediatenornicotine biosynthesis in Nicotiana tabacum L.: Functionalcharacterization of the CYP82E10 gene,” Phytochemistry, 71 (2010),1988-1998 (SEQ ID NO: 1, which sets forth a cyp82e4 W329Stop, herebyincorporated by reference in its entirety)), a mutated CYP82E5v2 recitedin Lewis et al. (supra) as 325-6 #1-13 (SEQ ID NO: 2, which sets forth acyp82e5v2 W422Stop, hereby incorporated by reference in its entirety),and a mutated CYP82E10 recited in Lewis et al. (supra) as 325-6 #1041(SEQ ID NO:11, which sets forth a cyp82e10 P381S, hereby incorporated byreference in its entirety). Mutations cyp82e4 W329Stop and cyp82e5v2W422Stop result in truncated proteins while cyp82E10 P381S results in anonfunctional protein. A cyp82e4 W329Stop (“e4”), a cyp82e5v2 W422Stop(“e5”), and a cyp82e10 P381S (“e10”) mutation are introduced from ae4e5|e4e5|e10e10 triple mutant from a strong converter burleybackground, line DH98-325-6, as listed in Table 2 of Lewis et al.(supra) into a flue-cured tobacco cultivar K326 background.

K326 SRC is the result of seven backcrosses with flue-cured cultivarK326 as the recurrent parent, followed by two rounds of selling withselection for homozygosity for the cyp82e4 W329Stop, the cyp82e5v2W422Stop, and cyp82e10 P381S mutations to yield BC₇F₃ plants (K326 SRC)in which the wild-type CYP82E4, CYP82E5v2 and CYP82E10 alleles of K326are replaced by the mutant (e.g., cyp82e4 W329Stop, cyp82e5v2 W422Stop,and cyp82e10 P381S) alleles.

K326 SRC progeny plants have genetic backgrounds that are at least 95%,at least 97%, at least 98%, or at least 99% similar to K326. K326 SRCplants exhibit low nornicotine levels and produce leaves with reducedpotential for accumulating derived NNN during curing, storage, andsmoking.

CMS K326 SRC

The present disclosure also provides tobacco cultivars, and partsthereof, from CMS K326 SRC, representative sample seeds of this cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-13454.The present disclosure also includes a tobacco plant, or part thereof,produced by growing a seed of CMS K326 SRC. A plant of the presentdisclosure can further include a plant with all, or essentially all, ofthe morphological and physiological characteristics of cultivar CMS K326SRC. CMS K326 SRC is a male-sterile (CMS) version of K326 SRC (CMSK326SRC) produced by crossing a plant of CMS K326 as a female withpollen of K326 SRC BC₆F₁ to produce male-sterile plants heterozygous forall three mutations. Progeny plants of the CMS K326×K326 SRC BC₆F₁ crossare male sterile. A plurality of CMS K326×K326 SRC BC₆F₁×CMS plants(e.g., CMS F₁ progeny plants) are screened for the cyp82e4 W329Stop,cyp82e5v2 W422Stop, and cyp82e10 P381S mutations to identify plantsheterozygous for all three mutations. A single male-sterile plantresulting from this cross is backcrossed as a female to K326 SRC toprepare BC₇F₁ CMS progeny. BC₇F₁ CMS progeny homozygous for the cyp82e4W329Stop, the cyp82e5v2 W422Stop, and cyp82e10 P381S mutations areidentified by genotyping and designated as CMS K326 SRC. Because the CMSK326 SRC line is male sterile, it is maintained via pollination withK326 SRC. K326 SRC is crossed as the male parent to CMS K326 SRC toprepare CMS K326 SRC F₁ progeny plants.

CMS K326 SRC and CMS K326 SRC F₁ progeny plants have genetic backgroundsthat are at least 95%, at least 97%, at least 98%, or at least 99%similar to K326. CMS K326 SRC and CMS K326 SRC F₁ progeny plants exhibitlow nornicotine levels and produce leaves with reduced potential foraccumulating derived NNN during curing, storage, and smoking.

K346 SRC

The present disclosure also provides tobacco cultivars, and partsthereof, from K346 SRC, where representative sample seeds of thiscultivar have been deposited with the ATCC under ATCC Accession No.PTA-13453. The present disclosure also includes a tobacco plant, or partthereof, produced by growing a seed of K346 SRC. A plant of the presentdisclosure can include a plant with all, or essentially all, of themorphological and physiological characteristics of cultivar K346 SRC.While not being limited by process, K346 SRC is a result of introducingthe cyp82e4 W329Stop, cyp82e5v2 W422Stop, and cyp82e10 P381S mutationsfrom DH98-325-6. F₁ individuals originating from a cross between K346and DH98-325-6 and heterozygous for each mutation are then backcrossedseven times to K346 to produce BC₇F₁ progeny. BC₇F₁ individualsheterozygous for all three mutations are self-pollinated to produceBC₇F₂ seed and individuals homozygous for all three mutationsidentified. A single BC₇F₂ plant is self-pollinated to produce BC₇F₃(K346 SRC) in which the wild-type CYP82E4, CYP82E5v2 and CYP82E10alleles of K346 are replaced by the mutant (e.g., cyp82e4 W329Stop,cyp82e5v2 W422Stop, and cyp82e10 P381S).

K346 SRC has a genetic background that is at least 95%, at least 97%, atleast 98%, or at least 99% similar to K346. K346 SRC exhibits lownornicotine levels and produce leaves with reduced potential foraccumulating derived NNN during curing, storage, and smoking.

CMS K346 SRC

The present disclosure also provides tobacco cultivars, and partsthereof, from CMS K346 SRC, representative sample seeds of this cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-13452.The present disclosure also includes a tobacco plant, or part thereof,produced by growing a seed of CMS K346 SRC. A plant of the presentdisclosure can further include a plant with all, or essentially all, ofthe morphological and physiological characteristics of cultivar CMS K426SRC. CMS K346 SRC is a male-sterile (CMS) version of K346 SRC (CMS K346SRC) produced by crossing a plant of CMS K346 as a female with pollen ofK346 SRC BC₆F₁ to produce male-sterile plants heterozygous for all threemutations. Progeny plants of the CMS K346×K346 SRC BC₆F₁ cross are malesterile. A plurality of CMS K346×K346 SRC BC₆F₁ CMS plants (e.g., CMS F₁progeny plants) are screened for the cyp82e4 W329Stop, cyp82e5v2W422Stop, and cyp82e10 P381S mutations to identify plants heterozygousfor all three mutations. A single male-sterile plant resulting from thiscross is backcrossed as a female to K346 SRC to prepare BC₇F₁ CMSprogeny. BC₇F₁ CMS progeny homozygous for the cyp82e4 W329Stop, thecyp82e5v2 W422Stop, and cyp82e10 P381S mutations are identified bygenotyping and designated as CMS K346 SRC. Because the CMS K346 SRC lineis male sterile, it is maintained via pollination with K346 SRC. K346SRC is crossed as the male parent to CMS K346 SRC to prepare CMS K346SRC F₁ progeny plants.

CMS K346 SRC and CMS K346 SRC F₁ progeny plants have genetic backgroundsthat are at least 95%, at least 97%, at least 98%, or at least 99%similar to K346. CMS K346 SRC and CMS K346 SRC F₁ progeny plants exhibitlow nornicotine levels and produce leaves with reduced potential foraccumulating derived NNN during curing, storage, and smoking.

NC1562-1 SRC

The present disclosure also provides tobacco cultivars, and partsthereof, from NC1562-1 SRC, where representative sample seeds of thiscultivar have been deposited with the ATCC under ATCC Accession No.PTA-13457. The present disclosure also includes a tobacco plant, or partthereof, produced by growing a seed of NC 1562-1 SRC. A plant of thepresent disclosure can include a plant with all, or essentially all, ofthe morphological and physiological characteristics of cultivar NC1562-1SRC. While not being limited by process, NC1562-1 SRC is a result ofintroducing the cyp82e4 W329Stop, cyp82e5v2 W422Stop, and cyp82e10 P381Smutations from DH98-325-6. F₁ individuals originating from a crossbetween NC1562-1 and DH98-325-6 and heterozygous for each mutation arethen backcrossed seven times to NC1562-1 to produce BC₇F₁ progeny. BC₇F₁individuals heterozygous for all three mutations are self-pollinated toproduce BC₇F₂ seed and individuals homozygous for all three mutationsidentified. A single BC₇F₂ plant is self-pollinated to produce BC₇F₃(NC1562-1 SRC) in which the wild-type CYP82E4, CYP82E5v2 and CYP82E10alleles of K346 are replaced by the mutant (e.g., cyp82e4 W329Stop,cyp82e5v2 W422Stop, and cyp82e10 P381S).

NC1562-1 SRC has a genetic background that is at least 95%, at least97%, at least 98%, or at least 99% similar to NC1562-1. NC1562-1 SRCexhibits low nornicotine levels and produce leaves with reducedpotential for accumulating derived NNN during curing, storage, andsmoking.

NCTG-61 SRC

The present disclosure also provides tobacco cultivars, and partsthereof, from NCTG-61 SRC, where representative sample seeds of thiscultivar have been deposited with the ATCC under ATCC Accession No.PTA-120314. The present disclosure also includes a tobacco plant, orpart thereof, produced by growing a seed of NCTG-61 SRC. A plant of thepresent disclosure can include a plant with all, or essentially all, ofthe morphological and physiological characteristics of cultivar NCTG-61SRC. While not being limited by process, NCTG-61 SRC is a result ofintroducing the cyp82e4 W329Stop, cyp82e5v2 W422Stop, and cyp82e10 P381Smutations from DH98-325-6. F₁ individuals originating from a crossbetween NCTG-61 and DH98-325-6 and heterozygous for each mutation arethen backcrossed seven times to NCTG-61 to produce BC₇F₁ progeny. BC₇F₁individuals heterozygous for all three mutations are self-pollinated toproduce BC₇F₂ seed and individuals homozygous for all three mutationsidentified. A single BC₇F₂ plant is self-pollinated to produce BC₇F₃(NCTG-61 SRC) in which the wild-type CYP82E4, CYP82E5v2 and CYP82E10alleles of K346 are replaced by the mutant (e.g., cyp82e4 W329Stop,cyp82e5v2 W422Stop, and cyp82e10 P381S).

NCTG-61 SRC has a genetic background that is at least 95%, at least 97%,at least 98%, or at least 99% similar to NCTG-61. NCTG-61 SRC exhibitslow nornicotine levels and produce leaves with reduced potential foraccumulating derived NNN during curing, storage, and smoking.

CMS NCTG-61 SRC

The present disclosure also provides tobacco cultivars, and partsthereof, from CMS NCTG-61 SRC, where representative sample seeds of thiscultivar have been deposited with the ATCC under ATCC Accession No.PTA-13456. The present disclosure also includes a tobacco plant, or partthereof, produced by growing a seed of CMS NCTG-61 SRC. A plant of thepresent disclosure can include a plant with all, or essentially all, ofthe morphological and physiological characteristics of cultivar CMSNCTG-61 SRC.

CMS NCTG-61 SRC is a male-sterile (CMS) version of NCTG-61 (CMS NCTG-61)produced by crossing a plant of CMS NCTG-61 as a female with pollen ofNCTG-61 SRC to produce male-sterile plants heterozygous for all threemutations. The CMS progeny plants of the CMS NCTG-61×NCTG-61 SRC BC₆F₁cross are male sterile. A plurality of CMS NCTG-61×NCTG-61 SRC BC₆F₁ CMSplants (e.g., CMS F₁ progeny plants) are screened for the cyp82e4W329Stop, cyp82e5v2 W422Stop, and cyp82e10 P381S mutations to identifyplants heterozygous for all three mutations. A single male-sterile plantresulting from this cross is backcrossed as a female to NCTG-61 SRC toprepare BC₇F₁ CMS progeny. BC₇F₁ CMS progeny homozygous for the cyp82e4W329Stop, the cyp82e5v2 W422Stop, and cyp82e10 P381S mutations areidentified by genotyping and designated as CMS NCTG-61 SRC. Because theCMS NCTG-61 SRC line is male sterile, it is maintained via pollinationwith NCTG-61 SRC. NCTG-61 SRC is crossed as the male parent to CMSNCTG-61 SRC to prepare CMS NCTG-61 SRC F₁ progeny plants.

CMS NCTG-61 SRC and CMS NCTG-61 SRC F₁ progeny plants have geneticbackgrounds that are at least 95%, at least 97%, at least 98%, or atleast 99% similar to NCTG-61. CMS NCTG-61 SRC and CMS NCTG-61 SRC F₁progeny plants exhibit low nornicotine levels and produce leaves withreduced potential for accumulating derived NNN during curing, storage,and smoking.

NC196 SRC

The present disclosure includes tobacco cultivars, and parts thereof,from NC196 SRC. In another aspect, the present disclosure includes atobacco plant, or part thereof, produced by growing the seed of NC196SRC. A plant of the present disclosure can include a plant with all, oressentially all, of the morphological and physiological characteristicsof cultivar NC196 SRC.

While not being limited by process, NC 196 SRC is produced bypollinating plants of CMS NCTG-61 SRC with pollen of NC1562-1 SRC.Again, not limited by any particular scientific theory, cyp82e4W329Stop, cyp82e5v2 W422Stop, and cyp82e10 P381S mutations all encodefor proteins with reduced or eliminated ability to convert nicotine ornornicotine. NC196 SRC has a genetic background that is at least 95%, atleast 97%, at least 98%, or at least 99% similar to flue-cured tobaccocultivar NC 196, a hybrid generated by pollinating plants of amale-sterile breeding line NCTG-61 with pollen produced by fertilebreeding line NC1562-1. NC 196 SRC exhibits low NNN and is not subjectto conversion to high NNN's.

Other Plants

The present disclosure includes a tobacco seed produced by crossing twoparent tobacco plants and harvesting the resultant tobacco seed, whereat least one parent tobacco plant is K326 SRC. In one aspect, the K326SRC is the male parent plant. In another aspect, the CMS K326 SRC is thefemale parent plant. One aspect of the present disclosure providestobacco plants that are homozygous at the cyp82e4, cyp82e5v2, andcyp82E10 loci for SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 11,respectively, and which share a genetic background that is greater than75%, 80%, 85%, 90%, 95%, 98%, or 99% similar to K326 or CMS K326. In oneaspect, approximately or greater than 50%, 75%, or 100% of a progeny'sgenetics is provided by a plant of the present disclosure that ishomozygous at the cyp82e4, cyp82e5v2, and cyp82E10 loci for SEQ ID NO:1, SEQ ID NO: 2, and SEQ ID NO: 11, respectively. In one aspect, a plantof the present disclosure has a genetic background that is at least 95%,at least 97%, at least 98%, or at least 99% similar to K326 or CMS K326.In another aspect, a plant of the present disclosure exhibits lownornicotine and is not subject to conversion to high NNN's. In oneaspect, a plant of the present disclosure is the progeny plant of afemale or male parent plant that is Fusarium wilt resistant. In anotheraspect, a plant of K326 SRC has low resistance to black shank andmoderate resistance to bacterial wilt.

The present disclosure includes a tobacco seed produced by crossing twoparent tobacco plants and harvesting the resultant tobacco seed, whereat least one parent tobacco plant is K346 SRC. In one aspect, the K346SRC is the male parent plant. In another aspect, the CMS K346 SRC is thefemale parent plant. One aspect of the present disclosure providestobacco plants that are homozygous at the cyp82e4, cyp82e5v2, andcyp82E10 loci for SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 11,respectively, and which share a genetic background that is greater than75%, 80%, 85%, 90%, 95%, 98%, or 99% similar to K346 or CMS K346. In oneaspect, approximately or greater than 50%, 75%, or 100% of a progeny'sgenetics is provided by a plant of the present disclosure that ishomozygous at the cyp82e4, cyp82e5v2, and cyp82E10 loci for SEQ ID NO:1, SEQ ID NO: 2, and SEQ ID NO: 11, respectively. In one aspect, a plantof the present disclosure has a genetic background that is at least 95%,at least 97%, at least 98%, or at least 99% similar to K346 or CMS K346.In another aspect, a plant of the present disclosure exhibits lownornicotine and is not subject to conversion to high NNN's. In oneaspect, a plant of the present disclosure is the progeny plant of afemale or male parent plant that is Fusarium wilt resistant. In anotheraspect, a plant of K346 SRC has high resistance to black shank and highresistance to bacterial wilt.

The present disclosure includes a tobacco seed produced by crossing twoparent tobacco plants and harvesting the resultant tobacco seed, whereat least one parent tobacco plant is NC1562-1 SRC. In one aspect, theNC1562-1 SRC is the male parent plant. In another aspect, the CMSNCTG-61 SRC is the female parent plant. One aspect of the presentdisclosure provides tobacco plants that are homozygous at the cyp82e4,cyp82e5v2, and cyp82E10 loci for SEQ ID NO: 1, SEQ ID NO: 2, and SEQ IDNO: 11, respectively, and which share a genetic background that isgreater than 75%, 80%, 85%, 90%, 95%, 98%, or 99% similar to NC1562-1 orCMS NC1562-1. In one aspect, approximately or greater than 50%, 75%, or100% of a progeny's genetics is provided by a plant of the presentdisclosure that is homozygous at the cyp82e4, cyp82e5v2, and cyp82E10loci for SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 11, respectively. Inone aspect, a plant of the present disclosure has a genetic backgroundthat is at least 95%, at least 97%, at least 98%, or at least 99%similar to NC1562-1 SRC and CMS NCTG-61 SRC. In another aspect, a plantof the present disclosure exhibits low nornicotine and is not subject toconversion to high NNN's. In one aspect, a plant of the presentdisclosure is the progeny plant of a female or male parent plant that isFusarium wilt resistant. In another aspect, a plant of NC196 SRC hasmoderate resistance to black shank and moderate resistance to bacterialwilt.

In one aspect, a plant of the present disclosure is a medium-latematuring variety with moderately high yield potential. In anotheraspect, a plant of the present disclosure offers a broad range ofimportant agronomic characteristics. In a further aspect, a plant of thepresent disclosure has one, two, three, four or more of the traitsincluding, but not limited to, moderate resistance to black shank, sometolerance to blue mold, black root rot resistance, and resistance tocommon virus diseases. In another aspect, a plant of the presentdisclosure has blue mold tolerance and level 4 resistance to both racesof black shank and high root rot resistance. In one aspect, a plant ofthe present disclosure, such as K326 SRC, CMS K326 SRC, K346 SRC, CMSK346 SRC, NC1562-1 SRC, NCTG-61 SRC, CMS NCTG-61 SRC, and hybridcultivar NC196 SRC, lacks Fusarium wilt resistance. In another aspect, aplant of the present disclosure is Fusarium wilt resistant. In anotheraspect, a plant of the present disclosure has low resistance to blackshank and moderate resistance to bacterial wilt.

In an aspect, the plants of the present disclosure have reduced oreliminated ability to convert nicotine to nornicotine. In an aspect, thepercentage nicotine conversion is less than 75%, 70%, 60%, 50%, or 25%of that found in K326, K346, NC1562-1, NCTG-61 or hybrid cultivar NC196.The nicotine conversion in plants of the present disclosure, including,but not limited to, K326 SRC, CMS K326 SRC, K346 SRC, CMS K346 SRC,NC1562-1 SRC, NCTG-61 SRC, CMS NCTG-61 SRC, and hybrid cultivar NC196SRC, can be less than about 4%, about 3.5%, about 3%, about 2.5%, about2%, about 1.5%, about 1%, about 3-1%, about 3-0.5%, or about 2-0.5%. Ina preferred aspect, the percentage nicotine conversion is less than 25%,10%, 5%, or 2% of that found in K326, K346, NC1562-1, NCTG-61 or hybridcultivar NC196 without the cyp82e4 W329Stop, the cyp82e5v2 W422Stop, andcyp82e10 P381S mutations. In an aspect, the tobacco plants of thepresent disclosure have a nicotine conversion rate of 3.5, 3.25, 3.0 or2.75% or less. In another aspect, the nicotine conversion rate oftobacco plants of the present disclosure can be 4.0, 3.9, 3.8, 3.7, 3.6,3.5, 3.4, 3.3, 3.2, 3.1, 3.0, 2.9, 2.8, 2.7, 2.6, 2.5, 2.4, 2.3, 2.2,2.1, 2.0, 1.9, 1.8, 1.7, 1.6, 1.5, 1.4, 1.3, 1.2, 1.1, 1.0, 0.9, 0.8,0.7, 0.6, 0.5% or less. In another aspect, the nicotine conversion rateof tobacco plants of the present disclosure can be 2.9, 2.8, 2.7, 2.6,2.5, 2.4, 2.3, 2.2, 2.1, 2.0, 1.9, 1.8, 1.7, 1.6, 1.5, 1.4, 1.3, 1.2,1.1, 1.0, 0.9, 0.8, 0.7, 0.6% or less. In another aspect, the nicotineconversion rates can be from 0.5 to 0.9%, 0.5 to 1.5%, 0.5 to 2.0%, 0.5to 2.5%, 0.5 to 2.75%, and 0.5 to 3.0%. In another aspect, the nicotineconversion rates can be from 1.0 to 1.5%, 1.0 to 1.75%, 1.0 to 2.0%, 1.0to 2.5%, 1.0 to 2.75%, and 1.0 to 3.0%. In another aspect, the nicotineconversion rate in a plant of the present disclosure may be less than2.9, 2.75, 2.5, 2.25, 2.0, 1.9, 1.8, 1.7, 1.6, 1.5, 1.4, 1.3, 1.2, 1.1or 1.0%. In an aspect, the tobacco plants of the present disclosure havea nicotine conversion rate of 3.5, 3.25, 3.0 or 2.75% or less. Inanother aspect, the nicotine conversion rate of tobacco plants of thepresent disclosure can be 2.9, 2.8, 2.7, 2.6, 2.5, 2.4, 2.3, 2.2, 2.1,2.0, 1.9, 1.8, 1.7, 1.6, 1.5, 1.4, 1.3, 1.2, 1.1, 1.0, 0.9, 0.8, 0.7,0.6% or less. In another aspect, the nicotine conversion rates can befrom 0.5 to 0.9%, 0.5 to 1.5%, 0.5 to 2.0%, 0.5 to 2.5%, 0.5 to 2.75%,and 0.5 to 3.0%. In another aspect, the nicotine conversion rates can befrom 1.0 to 1.5%, 1.0 to 1.75%, 1.0 to 2.0%, 1.0 to 2.5%, 1.0 to 2.75%,and 1.0 to 3.0%. In another aspect, the nicotine conversion rate in aplant of the present disclosure may be less than 2.9, 2.75, 2.5, 2.25,2.0, 1.9, 1.8, 1.7, 1.6, 1.5, 1.4, 1.3, 1.2, 1.1 or 1.0%.

In a further aspect, tobacco filler made from a plant of the presentdisclosure comprise a nicotine conversion rate between 0.05 and 0.5%,between 0.1 and 0.45%, between 0.15 and 0.4%, between 0.2 and 0.35%, orbetween 0.25 and 0.3%.

In a further aspect, tobacco filler made from a plant of the presentdisclosure comprise a nicotine conversion rate between 0.05 and 0.5%,between 0.1 and 0.5%, between 0.15 and 0.5%, between 0.2 and 0.5%,between 0.25 and 0.5%, between 0.3 and 0.5%, between 0.35 and 0.5%,between 0.4 and 0.5%, or between 0.45 and 0.5%.

In a further aspect, tobacco filler made from a plant of the presentdisclosure comprise a nicotine conversion rate between 0.05 and 0.5%,between 0.05 and 0.45%, between 0.05 and 0.4%, between 0.05 and 0.35%,between 0.05 and 0.3%, between 0.05 and 0.25%, between 0.05 and 0.2%,between 0.05 and 0.15%, or between 0.05 and 0.1%.

In another aspect, the tobacco plants of the present disclosuretypically have a reduced amount of nornicotine of less than about 0.10%dry weight. For example, the nornicotine content in such plants can be1.2, 1.0, 0.7, 0.5, 0.4, 0.2, 0.1, 0.09, 0.085, 0.08, 0.075, 0.07,0.065, 0.06, 0.055, 0.05, 0.045, 0.04, 0.035, 0.025, 0.01, 0.009,0.0075, 0.005, 0.0025, 0.001, 0.0009, 0.00075, 0.0005, 0.00025, or0.0001% dry weight, or undetectable. In another aspect, the nornicotinecontent can be less than 1.2, 1.0, 0.9, 0.8, 0.7, 0.5, 0.4, 0.2, 0.1,0.075, 0.05, 0.025, 0.01, 0.009, 0.0075, 0.005, 0.0025, 0.001, 0.0009,0.00075, 0.0005, 0.00025, or 0.0001% dry weight. In another aspect, thenornicotine content in such plants can be from 1.2-1.0, 0.7-0.5,0.4-0.2, 0.1-0.075, 0.05-0.025, 0.01-0.0075, 0.005-0.0025,0.001-0.00075, 0.0005-0.00025, or 0.0005-0.0001% dry weight. In a plantof the present disclosure, the nornicotine is a relatively smallpercentage of total alkaloids in the plant compared to a commercialseedlot of K326, K346, NC1562-1, NCTG-61 and hybrid cultivar NC196. Thenornicotine in a plant of the present disclosure may be 2-1%, less than3%, about 2%, about 1.5%, about 1%, or 0.75% of total alkaloids. Tobaccoproducts having a reduced amount of nitrosamine content can bemanufactured using tobacco plant material from plants and plant parts ofthe present disclosure. The tobacco product typically has a reducedamount of nornicotine of less than about 3 mg/g. For example, thenornicotine content in such a product can be 3.0 mg/g, 2.5 mg/g, 2.0mg/g, 1.5 mg/g, 1.0 mg/g, 750 μg/g, 500 pg/g, 250 pg/g, 100 pg/g, 75pg/g, 50 pg/g, 25 pg/g, 10 pg/g, 7.0 pg/g, 5.0 pg/g, 4.0 pg/g, 2.0 pg/g,1.0 pg/g, 0.5 pg/g, 0.4 pg/g, 0.2 pg/g, 0.1 pg/g, 0.05 pg/g, 0.01 pg/g,or undetectable. The tobacco product typically has a reduced amount ofNNN of less than about 10 pg/g. For example, the nornicotine content insuch a product can be about 10 pg/g, 7.0 pg/g, 5.0 pg/g, 4.0 pg/g, 2.0pg/g, 1.0 pg/g, 0.5 pg/g, 0.4 pg/g, 0.2 pg/g, 0.1 pg/g, 0.05 pg/g, 0.01pg/g, or undetectable. The percentage of secondary alkaloids relative tototal alkaloid content contained in a plant of the present disclosuremay not be statistically different than from a commercial seedlot ofK326, K346, NC1562-1, NCTG-61 and hybrid cultivar NC196.

In an aspect, a plant of the present disclosure or tobacco filler madetherefrom has reduced level of NNN. In an aspect, the NNN level is lessthan 75%, 70%, 60%, 50%, or 25% of that found in K326, K346, NC1562-1,NCTG-61 or hybrid cultivar NC196. The NNN level in tobacco filler madefrom plants of the present disclosure, including, but not limited to,K326 SRC, CMS K326 SRC, K346 SRC, CMS K346 SRC, NC1562-1 SRC, NCTG-61SRC, CMS NCTG-61 SRC, and hybrid cultivar NC196 SRC, can be less thanabout 0.5, 0.4, 0.3, 0.2, or 0.1 ppm. In a preferred aspect, the NNNlevel of a tobacco filler disclosed herein is between 0.8 and 0.05,between 0.8 and 0.1, between 0.8 and 0.2, between 0.8 and 0.3, between0.8 and 0.4, between 0.8 and 0.5, between 0.8 and 0.6, between 0.7 and0.05, between 0.6 and 0.05, between 0.5 and 0.05, between 0.4 and 0.05,between 0.3 and 0.05, between 0.2 and 0.05, between 0.1 and 0.05,between 0.7 and 0.1, between 0.6 and 0.2, or between 0.5 and 0.3 ppm. Ina preferred aspect, the NNN level is less than 25%, 10%, 5%, or 2% ofthat found in K326, K346, NC1562-1, NCTG-61 or hybrid cultivar NC196without the cyp82e4 W329Stop, the cyp82e5v2 W422Stop, and cyp82e10 P381Smutations.

In an aspect, a cigarette made from a plant of the present disclosurehas reduced level of NNN in its smoke. In an aspect, the cigarette smokeNNN level from plants of the instant disclosure is less than 75%, 70%,60%, 50%, or 25% of that found in K326, K346, NC1562-1, NCTG-61 orhybrid cultivar NC196. The cigarette smoke NNN level from plants of thepresent disclosure, including, but not limited to, K326 SRC, CMS K326SRC, K346 SRC, CMS K346 SRC, NC1562-1 SRC, NCTG-61 SRC, CMS NCTG-61 SRC,and hybrid cultivar NC196 SRC, can be less than about 0.035, 0.03,0.025, 0.02, 0.015, 0.01, or 0.005 ng/mg TPM. In a preferred aspect, thecigarette smoke NNN level is between 0.035 and 0.005, between 0.035 and0.01, between 0.035 and 0.015, between 0.035 and 0.02, between 0.035 and0.025, between 0.03 and 0.005, between 0.025 and 0.005, between 0.02 and0.005, between 0.015 and 0.005, between 0.03 and 0.01, or between 0.025and 0.015 ng/mg TPM. In a preferred aspect, the cigarette smoke NNNlevel is less than 25%, 10%, 5%, or 2% of that found in K326, K346,NC1562-1, NCTG-61 or hybrid cultivar NC196 without the cyp82e4 W329Stop,the cyp82e5v2 W422Stop, and cyp82e10 P381S mutations.

Differences between two inbred tobacco varieties or two hybrid tobaccovarieties can be evaluated using statistical approaches. Statisticalanalysis includes the calculation of mean values, determination of thestatistical significance of the sources of variation, and thecalculation of the appropriate variance components. Methods fordetermining statistical significance are known in the art. Statisticalsoftware is available, for example, the PROC GLM function of SAS.Significance is generally presented as a “p-value.” A statisticallysignificant p-value is less than 0.10. In a preferred aspect, thep-value is less than, or equal to, 0.05. In another aspect, the p-valueis 0.04 or less, 0.03 or less, or 0.02 or less. In yet another aspect, astatistically significant value is less than 0.01. In yet anotheraspect, it can be less than 0.009, less than 0.008, less than 0.007,less than 0.006, less than 0.005, less than 0.004, less than 0.003, lessthan 0.002, or less than 0.001.

Tobacco plants of the present disclosure that are homozygous for thecyp82e4 W329Stop, the cyp82e5v2 W422Stop, and cyp82e10 P381S alleleshave a reversion rate that is statistically significantly lower thancorresponding control low-converter plants having wild type nicotinedemethylase CYP82E4, E5, and E10 genes. In addition, homozygous CYP82E4,CYP82E5, and CYP82E10 triple mutant tobacco plants have a percentconversion to nornicotine of less than 2.0%, e.g., undetectable to 2.0%,1.0 to 2.0%, 0.8 to 1.8%, 0.8 to 2.0%, or 1.0 to 2.0%.

Nicotine and nornicotine can be measured in ethylene-treated leavesusing methods known in the art (e.g., gas chromatography). Percentnicotine demethylation in a sample is calculated by dividing the levelof nornicotine by the combined level of nicotine and nornicotine asmeasured in the sample, and multiplying by 100. Percent nicotinedemethylation in a sample from a plant of the present disclosure is 50,40, 30, 20, or 10 percent of a sample from an individual plant grownfrom a commercial seedlot of K326, K346, NC1562-1, NCTG-61 and hybridcultivar NC196.

In an aspect, the tobacco plants of the present disclosure have a USDAquality index of about 73, about 72, about 71, about 70, about 69, about68, about 67 or about 66. In an aspect, the tobacco plants of thepresent disclosure have a USDA quality index of about 65. In anotheraspect, the quality index may be at least about 55, 60, 62.5 or greater.In another aspect, tobacco plants of the present disclosure can have aquality index of 60-65, 60-70, 62.5-65, 62.5-70, or 65-70.

A plant of the present disclosure, including, but not limited to, K326SRC, CMS K326 SRC, K346 SRC, CMS K346 SRC, NC1562-1 SRC, NCTG-61 SRC,CMS NCTG-61 SRC, and hybrid cultivar NC196 SRC, can have any yield,including high (e.g., over 3000 lbs/A), moderately high (e.g., 2200-3000lbs/A), and moderate (e.g., less than 2000 lbs/A) yield potential.

In another aspect, the present disclosure also provides for a plantgrown from the seed of a K326 SRC, CMS K326 SRC, K346 SRC, CMS K346 SRC,NC1562-1 SRC, NCTG-61 SRC, CMS NCTG-61 SRC, or hybrid NC196 SRC plant inwhich alkaloids obtained from tobacco plants grown for the seed havedecreased nornicotine, as well as plant parts and tissue cultures fromsuch plants, representative sample seeds of these cultivars having beendeposited with the ATCC, for example, under ATCC Accession No. PTA-13455for K326 SRC, ATCC Accession No. PTA-13454 for CMS K326 SRC, ATCCAccession No. PTA-13453 for K346 SRC, ATCC Accession No. PTA-13452 forCMS K346 SRC, ATCC Accession No. PTA-13457 for NC1562-1 SRC, ATCCAccession No. PTA-120314 for NCTG-61 SRC, ATCC Accession No. PTA-13456for CMS NCTG-61 SRC, and ATCC Accession No. PTA-13458 for hybridcultivar NC196 SRC.

An aspect of the present disclosure provides for parts of the cultivarsK326 SRC, CMS K326 SRC, K346 SRC, CMS K346 SRC, NC1562-1 SRC, NCTG-61SRC, CMS NCTG-61 SRC, and hybrid cultivar NC196 SRC. For example,leaves, pollen, embryos, endosperm, cotyledons, hypocotyls, roots, roottips, anthers, flowers, ovules, shoots, stems, stalks, pith andcapsules, tissue culture comprising tissue, callus, cells or protoplastsof the cultivars K326 SRC, CMS K326 SRC, K346 SRC, CMS K346 SRC,NC1562-1 SRC, NCTG-61 SRC, CMS NCTG-61 SRC, and hybrid cultivar NC196SRC. In another aspect, the present disclosure provides for parts fromhybrids of cultivars K326 SRC, CMS K326 SRC, K346 SRC, CMS K346 SRC,NC1562-1 SRC, NCTG-61 SRC, CMS NCTG-61 SRC, and hybrid cultivar NC196SRC derived tobacco plants. In yet another aspect, the presentdisclosure provides for parts from genetically modified (e.g., byconventional breeding or genetic engineering techniques) forms of theforegoing plants and tissue culture.

In a further aspect, the present disclosure also provides for anon-regenerable cell of the cultivars K326 SRC, CMS K326 SRC, K346 SRC,CMS K346 SRC, NC1562-1 SRC, NCTG-61 SRC, CMS NCTG-61 SRC, and hybridcultivar NC196 SRC. In another aspect, the present disclosure alsoprovides for non-reproductive material or a non-reproductive cell of thecultivars K326 SRC, CMS K326 SRC, K346 SRC, CMS K346 SRC, NC1562-1 SRC,NCTG-61 SRC, CMS NCTG-61 SRC, and hybrid cultivar NC196 SRC.

Additional aspects of the present disclosure provide products comprisingtobacco wherein the tobacco further comprises tobacco from the plants ofthe present disclosure, and parts thereof. Other aspects of thedisclosure provide cured plant leaves and other plant parts.Accordingly, in some aspects, the cured plant parts include, but are notlimited to, a leaf, pollen, ovule, embryo, cotyledon, hypocotyl,meristematic cell, protoplast, root, root tip, pistil, anther, flower,shoot, stem, pod, petiole, and the like, and combinations thereof.

Thus, in some aspects, the present disclosure provides a cured tobaccocomprising the leaves of the tobacco plant designated K326 SRC, arepresentative sample seed of said cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13455. In another aspect, thepresent disclosure provides a cured tobacco comprising the leaves of thetobacco plant designated CMS K326 SRC, a representative sample seed ofsaid cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-13454. In another aspect, the present disclosure provides acured tobacco comprising the leaves of the tobacco plant designated K346SRC, a representative sample seed of said cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13453. In another aspect, thepresent disclosure provides a cured tobacco comprising the leaves of thetobacco plant designated CMS K346 SRC, a representative sample seed ofsaid cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-13452. In a further aspect, the present disclosure provides acured tobacco comprising the leaves of the tobacco plant designatedNC1562-1 SRC, a representative sample seed of said cultivar having beendeposited with the ATCC under ATCC Accession No, PTA-13457. In yetanother aspect, the present disclosure provides a cured tobaccocomprising the leaves of the tobacco plant designated NCTG-61 SRC, arepresentative sample seed of said cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-120314. In yet another aspect, thepresent disclosure provides a cured tobacco comprising the leaves of thetobacco plant designated CMS NCTG-61 SRC, a representative sample seedof said cultivar having been deposited with the ATCC under ATCCAccession No. PTA-13456. In yet another aspect, the present disclosureprovides a cured tobacco comprising the leaves of the hybrid tobaccoplant designated NC 196 SRC, a representative sample seed of said hybridcultivar having been deposited with the ATCC under ATCC Accession No.PTA-13458.

In an aspect, the present disclosure provides a cured tobacco comprisingthe stems of the tobacco plant designated K326 SRC, a representativesample seed of said cultivar having been deposited with the ATCC underATCC Accession No. PTA-13455. In another aspect, the present disclosureprovides a cured tobacco comprising the stems of the tobacco plantdesignated CMS K326 SRC, a representative sample seed of said cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-13454.In an aspect, the present disclosure provides a cured tobacco comprisingthe stems of the tobacco plant designated K346 SRC, a representativesample seed of said cultivar having been deposited with the ATCC underATCC Accession No. PTA-13453. In another aspect, the present disclosureprovides a cured tobacco comprising the stems of the tobacco plantdesignated CMS K346 SRC, a representative sample seed of said cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-13452.In a further aspect, the present disclosure provides a cured tobaccocomprising the stems of the tobacco plant designated NC1562-1 SRC, arepresentative sample seed of said cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13457. In yet another aspect, thepresent disclosure provides a cured tobacco comprising the stems of thetobacco plant designated NCTG-61 SRC, a representative sample seed ofsaid cultivar having been deposited with the ATCC under ATCC AccessionNo. PTA-120314. In yet another aspect, the present disclosure provides acured tobacco comprising the stems of the tobacco plant designated CMSNCTG-61 SRC, a representative sample seed of said cultivar having beendeposited with the ATCC under ATCC Accession No. PTA-13456. In yetanother aspect, the present disclosure provides a cured tobaccocomprising the stems of the hybrid tobacco plant designated NC 196 SRC,a representative sample seed of said hybrid cultivar having beendeposited with the ATCC under ATCC Accession No, PTA-13458.

In an aspect, the present disclosure provides a cured tobacco comprisingthe leaves and stems of the tobacco plants designated K326 SRC, arepresentative sample seed of said cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13455. In another aspect, thepresent disclosure provides a cured tobacco comprising the leaves andstems of the tobacco plants designated CMS K326 SRC, a representativesample seed of said cultivar having been deposited with the ATCC underATCC Accession No. PTA-13454. In an aspect, the present disclosureprovides a cured tobacco comprising the leaves and stems of the tobaccoplants designated K346 SRC, a representative sample seed of saidcultivar having been deposited with the ATCC under ATCC Accession No.PTA-13453. In another aspect, the present disclosure provides a curedtobacco comprising the leaves and sterns of the tobacco plantsdesignated CMS K346 SRC, a representative sample seed of said cultivarhaving been deposited with the ATCC under ATCC Accession No. PTA-13452.In a further aspect, the present disclosure provides a cured tobaccocomprising the leaves and stems of the tobacco plants designatedNC1562-1 SRC, a representative sample seed of said cultivar having beendeposited with the ATCC under ATCC Accession No, PTA-13457. In yetanother aspect, the present disclosure provides a cured tobaccocomprising the leaves and stems of the tobacco plants designated NCTG-61SRC, a representative sample seed of said cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-120314. In yet anotheraspect, the present disclosure provides a cured tobacco comprising theleaves and stems of the tobacco plants designated CMS NCTG-61 SRC, arepresentative sample seed of said cultivar having been deposited withthe ATCC under ATCC Accession No. PTA-13456. In yet another aspect, thepresent disclosure provides a cured tobacco comprising the leaves andstems of the hybrid tobacco plants designated NC196 SRC, arepresentative sample seed of said hybrid cultivar having been depositedwith the ATCC under ATCC Accession No. PTA-13458.

The present disclosure also provides a container of K326 SRC, CMS K326SRC, K346 SRC, CMS K346 SRC, NC1562-1 SRC, NCTG-61 SRC, CMS NCTG-61 SRC,or hybrid NC196 SRC seeds or other seeds of the present disclosure inwhich alkaloids obtained from tobacco plants grown from greater than 50%of the seeds have decreased nornicotine. In another aspect, alkaloidsobtained from K326 SRC, CMS K326 SRC, K346 SRC, CMS K346 SRC, NC1562-1SRC, NCTG-61 SRC, CMS NCTG-61 SRC, or hybrid NC196 SRC plants or otherplants of the present disclosure grown from greater than 55%, 60%, 65%,70%, 75%, 80%, 85%, 90%, or 95% of the seeds in the container havedecreased nornicotine, representative samples of seeds of thesecultivars having been deposited with the ATCC, for example, under ATCCAccession No. PTA-13455 for K326 SRC, ATCC Accession No. PTA-13454 forCMS K326 SRC, ATCC Accession No. PTA-13453 for K346 SRC, ATCC AccessionNo. PTA-13452 for CMS K346 SRC, ATCC Accession No. PTA-13457 forNC1562-1 SRC, ATCC Accession No. PTA-120314 for NCTG-61 SRC, ATCCAccession No. PTA-13456 for CMS NCTG-61 SRC, and ATCC Accession No.PTA-13458 for hybrid cultivar NC196 SRC.

The container of K326 SRC, CMS K326 SRC, K346 SRC, CMS K346 SRC,NC1562-1 SRC, NCTG-61 SRC, CMS NCTG-61 SRC, or hybrid NC196 SRC seeds orother seeds of the present disclosure may contain any number, weight orvolume of seeds. For example, a container can contain at least, orgreater than, about 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000,1500, 2000, 2500, 3000, 3500, 4000 or more seeds. Alternatively, thecontainer can contain at least, or greater than, about 1 ounce, 5ounces, 10 ounces, 1 pound, 2 pounds, 3 pounds, 4 pounds, 5 pounds ormore seeds. Representative samples of seeds of these cultivars havingbeen deposited with the ATCC, for example, under ATCC Accession No.PTA-13455 for K326 SRC, ATCC Accession No. PTA-13454 for CMS K326 SRC,ATCC Accession No. PTA-13453 for K346 SRC, ATCC Accession No. PTA-13452for CMS K346 SRC, ATCC Accession No. PTA-13457 for NC1562-1 SRC, ATCCAccession No. PTA-120314 for NCTG-61 SRC, ATCC Accession No. PTA-13456for CMS NCTG-61 SRC, and ATCC Accession No. PTA-13458 for hybridcultivar NC196 SRC.

Containers of K326 SRC, CMS K326 SRC, K346 SRC, CMS K346 SRC, NC1562-1SRC, NCTG-61 SRC, CMS NCTG-61 SRC, or hybrid NC196 SRC seeds or otherseeds of the present disclosure may be any container available in theart. By way of a non-limiting example, a container may be a box, a bag,a packet, a pouch, a tape roll, a pail, a foil, a tube, or a bottle.Representative samples of seeds of these cultivars having been depositedwith the ATCC, for example, under ATCC Accession No. PTA-13455 for K326SRC, ATCC Accession No, PTA-13454 for CMS K326 SRC, ATCC Accession No.PTA-13453 for K346 SRC, ATCC Accession No. PTA-13452 for CMS K346 SRC,ATCC Accession No. PTA-13457 for NC1562-1 SRC, ATCC Accession No.PTA-120314 for NCTG-61 SRC, ATCC Accession No, PTA-13456 for CMS NCTG-61SRC, and ATCC Accession No. PTA-13458 for hybrid cultivar NC196 SRC.

In another aspect, the present disclosure also provides a container ofK326 SRC, CMS K326 SRC, K346 SRC, CMS K346 SRC, NC1562-1 SRC, NCTG-61SRC, CMS NCTG-61 SRC, or hybrid cultivar NC196 SRC in which greater than50% of K326 SRC, CMS K326 SRC, K346 SRC, CMS K346 SRC, NC1562-1 SRC, CMSNCTG-61 SRC, or hybrid NC196 SRC seeds or other seeds of the presentdisclosure have decreased nornicotine. Representative samples of seedsof these cultivars having been deposited with the ATCC, for exampleunder ATCC Accession No. PTA-13455 for K326 SRC, ATCC Accession No.PTA-13454 for CMS K326 SRC, ATCC Accession No. PTA-13453 for K346 SRC,ATCC Accession No. PTA-13452 for CMS K346 SRC, ATCC Accession No.PTA-13457 for NC1562-1 SRC, ATCC Accession No. PTA-120314 for NCTG-61SRC, ATCC Accession No. PTA-13456 for CMS NCTG-61 SRC, and ATCCAccession No. PTA-13458 for hybrid cultivar NC196 SRC.

In one aspect, the present disclosure provides a seed of a K326 SRC, CMSK326 SRC, K346 SRC, CMS K346 SRC, NC1562-1 SRC, NCTG-61 SRC, CMS NCTG-61SRC, or hybrid NC196 SRC plant or other plant of the present disclosurein which a plant grown from a seed is male sterile. Representativesamples of seeds of these cultivars having been deposited with the ATCC,for example, under ATCC Accession No. PTA-13455 for K326 SRC, ATCCAccession No. PTA-13454 for CMS K326 SRC, ATCC Accession No. PTA-13453for K346 SRC, ATCC Accession No. PTA-13452 for CMS K346 SRC, ATCCAccession No. PTA-13457 for NC1562-1 SRC, ATCC Accession No. PTA-120314for NCTG-61 SRC, ATCC Accession No. PTA-13456 for CMS NCTG-61 SRC, andATCC Accession No. PTA-13458 for hybrid cultivar NC196 SRC.

The instant disclosure further provides cured tobacco material, tobaccoblends, and tobacco products made from tobacco plants disclosed herein.In one aspect, the cured tobacco material of the instant disclosure isflue-cured. In another aspect, the cured tobacco material of the instantdisclosure is sun-cured, air-cured, or fire-cured.

Tobacco material obtained from the tobacco lines, varieties or hybridsof the present disclosure can be used to make tobacco productsincluding, without limitation, cigarette products (e.g., cigarettes andbidi cigarettes), cigar products (e.g., cigar wrapping tobacco andcigarillos), pipe tobacco products, products derived from tobacco,tobacco derived nicotine products, smokeless tobacco products (e.g.,moist snuff, dry snuff, and chewing tobacco), films, chewables, tabs,shaped parts, gels, consumable units, insoluble matrices, hollow shapesand the like. See, e.g., U.S. Patent Publication No. US 2006/0191548,which is herein incorporated by reference in its entirety.

Tobacco products derived from plants of the present disclosure alsoinclude cigarettes and other smoking articles, particularly thosesmoking articles including filter elements, wherein the rod of smokeablematerial includes cured tobacco within a tobacco blend. In an aspect, atobacco product may be pipe tobacco, cigar tobacco, cigarette tobacco,chewing tobacco, leaf tobacco, shredded tobacco, and cut tobacco.

In an aspect, a tobacco product of the instant disclosure is selectedfrom the group consisting of a cigarillo, a non-ventilated recess filtercigarette, a vented recess filter cigarette, a cigar, snuff, pipetobacco, cigar tobacco, cigarette tobacco, chewing tobacco, leaftobacco, hookah tobacco, shredded tobacco, and cut tobacco. In anotheraspect, a tobacco product of the instant disclosure is a smokelesstobacco product. In a further aspect, a tobacco product of the instantdisclosure is selected from the group consisting of loose leaf chewingtobacco, plug chewing tobacco, moist snuff, and nasal snuff. In yetanother aspect, a tobacco product of the instant disclosure is selectedfrom the group consisting of an electronically heated cigarette, ane-cigarette, an electronic vaporing device.

In an aspect, the tobacco product of the present disclosure can be ablended tobacco product. In other aspects of the disclosure, the tobaccoproduct of the present disclosure can be a reduced nicotine tobaccoproduct. In still other aspects, the tobacco product of the presentdisclosure can be a blended tobacco product with reduced nicotinecontent. Thus, the tobacco product of the present disclosure can be ablended reduced nicotine tobacco product. Tobacco product materialcomprises a blend of tobacco materials from the present disclosure,wherein the blend comprises at least about 5, 10, 20, 30, 40, 50, 60,70, 80, 90, or 95 percent by weight of a cured tobacco, based on the dryweight of the tobacco material. US 2008/0245377 is herein incorporatedby reference for blend mixtures in its entirety.

In an aspect, tobacco products having a reduced amount of nitrosaminecontent can be manufactured using tobacco plant material from plants andplant parts of the present disclosure. The tobacco product typically hasa reduced amount of nornicotine of less than about 3 mg/g. For example,the nornicotine content in such a product can be 3.0 mg/g, 2.5 mg/g, 2.0mg/g, 1.5 mg/g, 1.0 mg/g, 750 μg/g, 500 pg/g, 250 pg/g, 100 pg/g, 75pg/g, 50 pg/g, 25 pg/g, 10 pg/g, 7.0 pg/g, 5.0 pg/g, 4.0 pg/g, 2.0 pg/g,1.0 pg/g, 0.5 pg/g, 0.4 pg/g, 0.2 pg/g, 0.1 pg/g, 0.05 pg/g, 0.01 pg/g,or undetectable. The tobacco product typically has a reduced amount ofNNN of less than about 10 pg/g. For example, the nornicotine content insuch a product can be about 10 pg/g, 7.0 pg/g, 5.0 pg/g, 4.0 pg/g, 2.0pg/g, 1.0 pg/g, 0.5 pg/g, 0.4 pg/g, 0.2 pg/g, 0.1 pg/g, 0.05 pg/g, 0.01pg/g, or undetectable. The percentage of secondary alkaloids relative tototal alkaloid content contained in a plant of the present disclosuremay not be statistically different than from a commercial seedlot ofK326, K346, NC1562-1, NCTG-61 and hybrid cultivar NC196.

The instant disclosure further provides a method manufacturing a tobaccoproduct, where the method comprising conditioning aged tobacco materialmade from an tobacco plant disclosed herein to increase its moisturecontent from between about 12.5% and about 13.5% to about 21%, blendingthe conditioned tobacco material to produce a desirable blend. In oneaspect, the method of manufacturing a tobacco product disclosed hereinfurther comprises casing or flavoring the blend. Generally, during thecasing process, casing or sauce materials are added to blends to enhancetheir quality by balancing the chemical composition and to developcertain desired flavor characteristics. Further details for the casingprocess can be found in Tobacco Production, Chemistry and Technology,Edited by L. Davis and M. Nielsen, Blackwell Science, 1999.

A tobacco plant of the present disclosure designated K326 SRC, CMS K326SRC, K346 SRC, CMS K346 SRC, NC1562-1 SRC, NCTG-61 SRC, CMS NCTG-61 SRC,or hybrid cultivar NC196 SRC, carrying the cyp82e4 W329Stop, thecyp82e5v2 W422Stop, and cyp82e10 P381S alleles can be used in a plantbreeding program to create useful lines, cultivars, varieties, progeny,inbreds, and hybrids. Thus, in some aspects, an F₁, F₂, F₃, or latergeneration tobacco plant containing the cyp82e4 W329Stop, the cyp82e5v2W422Stop, and cyp82e10 P381S alleles is crossed with a second Nicotianaplant, and progeny of the cross are identified in which the cyp82e4W329Stop, the cyp82e5v2 W422Stop, and cyp82e10 P381S alleles arepresent. It will be appreciated that the second Nicotiana plant will beK326, K346, NC1562-1, NCTG-61 and hybrid cultivar NC196 or any otherNicotiana species or line, optionally with an additional desirabletrait, such as herbicide resistance mentioned below.

In still other aspects, methods of the present disclosure furtherinclude self-pollinating or pollinating a male sterile pollen acceptorwith a pollen donor capable of being used in production of a progeny ofthe present disclosure, such as a male sterile hybrid of the presentdisclosure. Either the male sterile pollen acceptor plant or the pollendonor plant has at least one mutant allele, two, or even three mutantalleles at a nicotine demethylase locus, such as the cyp82e4 W329Stop,the cyp82e5v2 W422Stop, and cyp82e10 P381S. In an aspect, all threealleles at each nicotine demethylase locus are mutant alleles, makingthe plant homozygous for cyp82e4 W329Stop, the cyp82e5v2 W422Stop, andcyp82e10 P381S.

Breeding can be carried out via any known procedures. DNAfingerprinting, SNP or similar technologies may be used in amarker-assisted selection (MAS) breeding program to transfer or breedmutant alleles of a nicotine demethylase gene into other tobaccos. Forexample, a breeder can create segregating populations fromhybridizations of a genotype containing cyp82e4 W329Stop, the cyp82e5v2W422Stop, and cyp82e10 P381S alleles with an agronomically desirablegenotype. Plants in the F₂ or backcross generations can be screenedusing a marker developed from cyp82e4 W329Stop, the cyp82e5v2 W422Stop,or cyp82e10 P381S alleles or a fragment thereof, using one of thetechniques known in the art or listed herein. Plants identified aspossessing one or more cyp82e4 W329Stop, the cyp82e5v2 W422Stop, andcyp82e10 P381S alleles can be backcrossed or self-pollinated to create asecond population to be screened. Depending on the expected inheritancepattern or the MAS technology used, it may be necessary toself-pollinate the selected plants before each cycle of backcrossing toaid identification of the desired individual plants. Backcrossing orother breeding procedure can be repeated until the desired phenotype ofthe recurrent parent is recovered. A recurrent parent in the presentdisclosure can be K326 SRC, CMS K326 SRC, K346 SRC, CMS K346 SRC,NC1562-1 SRC, NCTG-61 SRC, CMS NCTG-61 SRC, or NC 196 SRC. Otherbreeding techniques can be found, for example, in Wernsman, E. A., andRufty, R. C. 1987. Chapter Seventeen. Tobacco. Pages 669-698 In:Cultivar Development. Crop Species. W. H. Fehr (ed.), MacMillanPublishing Go., Inc., New York, N.Y., incorporated herein by referencein their entirety.

Nicotiana species which exhibit breeding compatibility with Nicotianatabacum include Nicotiana amplexicaulis, PI 271989; Nicotianabenthamiana PI 555478; Nicotiana bigelovii PI 555485; Nicotiana debneyi;Nicotiana excelsior PI 224063; Nicotiana glutinosa PI 555507; Nicotianagoodspeedii PI 241012; Nicotiana gossei PI 230953; Nicotiana hesperis PI271991; Nicotiana knightiana PI 555527; Nicotiana maritima PI 555535;Nicotiana megalosiphon PI 555536; Nicotiana nudicaulis PI 555540;Nicotiana paniculata PI 555545; Nicotiana plumbaginifolia PI 555548;Nicotiana repanda PI 555552; Nicotiana rustica; Nicotiana suaveolens PI230960; Nicotiana sylvestris PI 555569; Nicotiana tomentosa PI 266379;Nicotiana tomentosiformis; and Nicotiana trigonophylla PI 555572. Seealso, Compendium of Tobacco Diseases published by AmericanPhytopathology Society, or The Genus Nicotiana Illustrated, published byJapan Tobacco Inc, hereby incorporated by reference in their entirety.

The result of a plant breeding program using the mutant tobacco plantsdescribed herein includes useful lines, cultivars, varieties, progeny,inbreds, and hybrids. As used herein, the term “cultivar” or “variety”refers to a population of plants that share constant characteristicswhich separate them from other plants of the same species. A cultivar orvariety is often, although not always, sold commercially. Whilepossessing one or more distinctive traits, a cultivar or variety isfurther characterized by a very small overall variation betweenindividuals within that variety. A “pure line” variety may be created byseveral generations of self-pollination and selection, or vegetativepropagation from a single parent using tissue or cell culturetechniques. A cultivar or variety can be essentially derived fromanother cultivar, line, or variety. As defined by the InternationalConvention for the Protection of New Varieties of Plants (Dec. 2, 1961,as revised at Geneva on Nov. 10, 1972, on Oct. 23, 1978, and on Mar. 19,1991), a cultivar or variety is “essentially derived” from an initialcultivar or variety if: a) it is predominantly derived from the initialcultivar or variety, or from a cultivar or variety that is predominantlyderived from the initial cultivar or variety, while retaining theexpression of the essential characteristics that result from thegenotype or combination of genotypes of the initial cultivar or variety;b) it is clearly distinguishable from the initial cultivar or variety;and c) except for the differences which result from the act ofderivation, it conforms to the initial cultivar or variety in theexpression of the essential characteristics that result from thegenotype or combination of genotypes of the initial cultivar or variety.Essentially derived varieties can be obtained, for example, by theselection of a natural or induced mutant, a somaclonal variant, avariant individual from plants of the initial variety, backerossing, ortransformation. A “line” as distinguished from a cultivar or varietymost often denotes a group of plants used non-commercially, for examplein plant research. A line typically displays little overall variationbetween individuals for one or more traits of interest, although theremay be some variation between individuals for other traits.

Hybrid tobacco varieties can be produced by preventing self-pollinationof female parent plants (i.e., seed parents) of a first variety,permitting pollen from male parent plants of a second variety tofertilize the female parent plants, and allowing F₁ hybrid seeds to formon the female plants. Self-pollination of female plants can be preventedby emasculating the flowers at an early stage of flower development.Alternatively, pollen formation can be prevented on the female parentplants using a form of male sterility. For example, male sterility canbe produced by cytoplasmic male sterility (CMS), or transgenic malesterility wherein a transgene inhibits microsporogenesis and/or pollenformation, or self-incompatibility. Female parent plants containing CMSare particularly useful. In aspects in which the female parent plantsare CMS, pollen may be harvested from male fertile plants and appliedmanually to the stigmas of CMS female parent plants, and the resultingF₁ seed is harvested.

Plants can be used to form single-cross tobacco F₁ hybrids.Alternatively, three-way crosses can be carried out wherein asingle-cross F₁ hybrid is used as a female parent and is crossed with adifferent male parent. As another alternative, double-cross hybrids canbe created wherein the F₁ progeny of two different single-crosses arethemselves crossed. Self-incompatibility can be used to particularadvantage to prevent self-pollination of female parents when forming adouble-cross hybrid.

Successful crosses yield F₁ plants that are fertile, have cyp82e4W329Stop, the cyp82e5v2 W422Stop, and cyp82e10 P381S alleles, and can bebackcrossed with one of the parents, such as K326 SRC, CMS K326 SRC,K346 SRC, CMS K346 SRC, NC1562-1 SRC, NCTG-61 SRC, CMS NCTG-61 SRC, orNC 196 SRC, if desired. In some aspects, a plant population in the F₂generation is screened for cyp82e4 W329Stop, the cyp82e5v2 W422Stop, andcyp82e10 P381S alleles. Selected plants can be crossed with one of theparents and the first backcross (BC1) generation plants areself-pollinated to produce a BC1 F₂ population that is again screenedfor variant nicotine demethylase gene expression (e.g., the null versionof the nicotine demethylase gene). The process of backcrossing,self-pollination, and screening is repeated, for example, at least 4times, until the final screening produces a plant that is fertile andreasonably similar to the recurrent parent. This plant, if desired, isself-pollinated and the progeny are subsequently screened again toconfirm that the plant exhibits the same low nicotine conversionphenotype as K326 SRC, CMS K326 SRC, K346 SRC, CMS K346 SRC, NC1562-1SRC, NCTG-61 SRC, CMS NCTG-61 SRC, or NC 196 SRC. Breeder's seed of theselected plant is produced using standard methods including, forexample, field testing, confirmation of the null condition for nicotinedemethylase, chemical analyses of cured leaf to determine the level ofalkaloids and/or chemical analyses of cured leaf to determine the ratioof nornicotine to nicotine+nornicotine.

In one aspect, a F₁ progeny is the result of a cross between K326 SRCand CMS K326 SRC to generate F₁ progeny that are male sterile. Inanother aspect, a F₁ progeny is the result of a cross between K346 SRCand CMS K346 SRC to generate F₁ progeny that are male sterile. Malesterile tobacco plants may be produced by any method known in the art.Methods of producing male sterile tobacco are described in Wernsman, E.A., and Rufty, R. C. 1987. Chapter Seventeen. Tobacco. Pages 669-698 In:Cultivar Development. Crop Species. W. H. Fehr (ed.), MacMillanPublishing Go., Inc., New York, N.Y. 761 pp.

The present disclosure further provides methods of producing a tobaccoplant by crossing one of cultivars K326 SRC, CMS K326 SRC, K346 SRC, CMSK346 SRC, NC1562-1 SRC, NCTG-61 SRC, CMS NCTG-61 SRC, and hybrid NC196SRC with itself or a different tobacco line. The disclosure furtherrelates to methods for producing other tobacco cultivars or breedinglines derived from cultivars K326 SRC, CMS K326 SRC, K346 SRC, CMS K346SRC, NC1562-1 SRC, NCTG-61 SRC, CMS NCTG-61 SRC, and hybrid NC196 SRC bycrossing a plant of cultivars K326 SRC, CMS K326 SRC, K346 SRC, CMS K346SRC, NC1562-1 SRC, CMS NCTG-61 SRC, and hybrid NC196 SRC with a secondtobacco plant and growing the progeny seed to yield a K326 SRC, CMS K326SRC, K346 SRC, CMS K346 SRC, NC1562-1 SRC, NCTG-61 SRC, CMS NCTG-61 SRC,or hybrid NC196 SRC-derived tobacco plant. An additional aspect of thepresent disclosure provides a method for a tobacco plant that containsin its genetic material one or more transgenes, comprising crossingcultivars of the present disclosure with a second cultivar containingone or more transgenes wherein progeny are produced, so that the geneticmaterial of the progeny that result from the cross comprise thetransgene(s) optionally operably linked to one or more regulatoryelements. In one aspect, the second cultivar may be a plant derived fromcultivars K326 SRC, CMS K326 SRC, K346 SRC, CMS K346 SRC, NC1562-1 SRC,NCTG-61 SRC, CMS NCTG-61 SRC, or hybrid NC196 SRC transformed with oneor more transgenes.

The disclosure further provides for the vegetative propagation of aplant of cultivars K326 SRC, CMS K326 SRC, K346 SRC, CMS K346 SRC,NC1562-1 SRC, NCTG-61 SRC, CMS NCTG-61 SRC, and hybrid NC196 SRC,hybrids and progeny thereof. In one aspect, the disclosure provides fora method of vegetatively propagating a plant of a tobacco cultivarcomprising collecting tissue capable of being propagated from a plant ofcultivars K326 SRC, CMS K326 SRC, K346 SRC, CMS K346 SRC, NC1562-1 SRC,NCTG-61 SRC, CMS NCTG-61 SRC, and hybrid NC196 SRC, cultivating thetissue to obtain a proliferated shoot and rooting the proliferatedshoots to obtain a rooted plantlet. In another aspect, the plant tissuemay be collected from an F₁ hybrid of a plant of cultivars K326 SRC, CMSK326 SRC, K346 SRC, CMS K346 SRC, NC1562-1 SRC, NCTG-61 SRC, CMS NCTG-61SRC, and hybrid NC196 SRC. In an aspect, the plant tissue may becollected from an F₂, F₃, F₄ or later progeny plant obtained by breedinga plant of cultivars K326 SRC, CMS K326 SRC, K346 SRC, CMS K346 SRC,NC1562-1 SRC, NCTG-61 SRC, CMS NCTG-61 SRC, and hybrid NC196 SRC.

A plant comprising a mutation in a nicotine demethylase gene can beidentified by selecting or screening the mutagenized plant material, orprogeny thereof. Such screening and selection methodologies are known tothose having ordinary skill in the art. Examples of screening andselection methodologies include, but are not limited to, Southernanalysis, PCR amplification for detection of a polynucleotide, Northernblots, RNase protection, primer-extension, RT-PCR amplification fordetecting RNA transcripts, enzymatic assays for detecting enzyme orribozyme activity of polypeptides and polynucleotides, and protein gelelectrophoresis, Western blots, immunoprecipitation, and enzyme-linkedimmunoassays to detect polypeptides. Other techniques such as in situhybridization, enzyme staining, and immunostaining also can be used todetect the presence or expression of polypeptides and/orpolynucleotides. Methods for performing all of the referenced techniquesare known.

It is understood that a tobacco plant of the present disclosure,including, but not limited to, K326 SRC, CMS K326 SRC, K346 SRC, CMSK346 SRC, NC1562-1 SRC, NCTG-61 SRC, CMS NCTG-61 SRC, and hybridcultivar NC196 SRC, can be transformed by a genetic construct ortransgene using a technique known in the art. Without limitation, anexample of a desired trait is herbicide resistance, pest resistance,disease resistance, high yield, high grade index, curability, curingquality, mechanical harvestability, holding ability, leaf quality,height, plant maturation (e.g., early maturing, early to mediummaturing, medium maturing, medium to late maturing, or late maturing),stalk size (e.g., small, medium, or large stalk), or leaf number perplant (e.g., small (e.g., 5-10 leaves), medium (e.g., 11-15 leaves), orlarge (e.g., 16-21) number of leaves), or any combination. Any plant ofthe present disclosure can be used as a basis for tissue culture,regenerated, transformed, or a combination of any of these. In anaspect, a plant of the present disclosure derived by tissue culture,transformation, or both has all of the morphological and physiologicalcharacteristics of cultivar K326 SRC, CMS K326 SRC, K346 SRC, CMS K346SRC, NC1562-1 SRC, CMS NCTG-61 SRC, or hybrid NC196 SRC.

Having now generally described the disclosure, the same will be morereadily understood through reference to the following examples that areprovided by way of illustration, and are not intended to be limiting ofthe present disclosure, unless specified.

EXAMPLES Example 1 Breeding of Homozygous cyp82e4 W329Stop, thecyp82e5v2 W422Stop, and cyp82e10 P381S Mutant Plants into the K326Flue-Cured Tobacco Cultivar

K326 SRC is a backcross-derived version of flue-cured tobacco cultivarK326 carrying introduced mutations in three genes (CYP82E4 (SEQ ID NO:5), CYP82E5 (SEQ ID NO: 8), and CYP82E10 (SEQ ID NO: 9)) previouslydocumented to encode for nicotine demethylase enzymes (Lewis et al.,2010). The introduced mutations in CYP82E4 (cyp82e4 W329Stop (SEQ ID NO:1)) and CYP82E5 (cyp82e5v2 W422Stop (SEQ ID NO: 2)) encode for prematurestop codons which render the genes non-functional. The introducedmutation in CYP82E10 (cyp82e10 P381S (SEQ ID NO: 11)) does not encodefor a premature stop codon, but does render the gene productnon-functional for converting nicotine to nornicotine (Lewis et al.2010). When in homozygous condition, the three mutations result intobacco plants with (1) reduced genetic capacity to demethylate nicotineto form nornicotine, and (2) a corresponding diminished potential toaccumulate N-nitrosonornicotine (NNN), a potent carcinogen found in manytobacco products. The mutation in CYP82E4 (cyp82e4 W329Stop (SEQ ID NO:1)) also provides phenotypic stability for the “nicotine conversion”trait and eliminates the requirement to utilize the cumbersome “LC”method for reducing levels of nornicotine in tobacco cultivars.

The original tobacco cultivar K326 is a fertile inbred line. CMS K326 isa cytoplasmic male-sterile version of K326. To develop K326 SRC, anindividual plant of K326 is pollinated with a plant of the geneticbackground DH98-325-6 carrying mutations in each of the three nicotinedemethylase genes. F₁ individuals originating from this cross andheterozygous for each mutation are backcrossed to the recurrent parent,K326, to produce BC₁F₁ progeny. BC₁F₁ progeny are screened usinggenotyping methodologies to identify individuals heterozygous for allthree mutations. A single triple heterozygous BC₁F₁ plant is backcrossedto K326 to produce BC₂F₁ progeny. The process of backcrossing andidentification of individuals heterozygous for all three mutations isrepeated through the BC₃F₁, BC₄F₁, BC₅F₁, BC₆F₁, and BC₇F₁ stages. Atthe BC₇F₁ stage, individuals heterozygous for all three mutations fromeach pedigree are self-pollinated to produce BC₇F₂ seed. A large numberof BC₇F₂ progeny from each pedigree are genotyped to identifyindividuals homozygous for all three mutations. A single BC₇F₂ planthomozygous for all three mutations is self-pollinated to produce a BC₇F₃family (K326 SRC) in which the wild-type CYP82E4, CYP82E5v2, andCYP82E10 alleles of K326 were replaced by the mutant (cyp82e4 W329Stop,cyp82e5v2 W422Stop, and cyp82e10 P381S alleles).

The male-sterile (CMS) version of K326 SRC (CMS K326 SRC) is produced bycrossing a plant of CMS K326 as a female with pollen of K326 SRC toproduce male-sterile plants heterozygous for all three mutations. Asingle male-sterile plant resulting from this cross is then subsequentlybackcrossed as a female to K326 SRC to produce progeny that aresegregating for individuals homozygous for all three mutations. Triplehomozygous individuals are identified by DNA genotyping to produce theCMS K326 SRC line. Because the line is male-sterile, it is maintainedvia pollination with K326 SRC.

Commercial K326 SRC is produced by pollinating plants of CMS K326 SRCwith pollen of K326 SRC.

Testing of K 326 SRC

K326 SRC is evaluated for cured leaf chemistry, yield, and physicalquality at three North Carolina field research locations during 2012(Oxford, Rocky Mount, Kinston). K326 is included for comparison. Theexperimental design at each location is a randomized complete blockdesign with four replications. Experimental units are single 20-plantplots. Plots are harvested and flue-cured. Plot weights are used todetermine per acre yields. Cured leaf is evaluated by a former USDAtobacco grader. Fifty gram composite leaf samples are collected fromeach plot and analyzed for percent nicotine, nornicotine, anatabine,anabasine, and percent nicotine conversion using gas chromatographyequipment.

Comparisons using the least significant difference (LSD) test indicatesthat K326 SRC has significantly (P<0.05) lower levels of nornicotine andpercent nicotine conversion relative to K326 (Table 1). K326 and K326SRC are not significantly different from each other for percentnicotine, yield, or cured leaf quality indices.

Analysis of NNN Levels in K 326 SRC Tobacco Filler and Cigarette Smoke

In an independent testing, K326 SRC is further evaluated for NNN levelsin both tobacco filler and cigarette smoke by gas chromatography.Percent nicotine conversion in tobacco filler is also measured. K326 isincluded for comparison. K326 SRC has a lower level of NNN compared toK326 in both tobacco filler (˜022 versus ˜0.53 ppm) and cigarette smoke(˜0.015 versus ˜0.04 ng/mg Total Particle Matter (TPM)) (FIG. 1).Percent nicotine conversion is lower in K326 SRC tobacco filler (˜0.8%)than K326 tobacco filler (˜3.1%) (FIG. 1).

Example 2 Breeding of Homozygous cyp82e4 W329Stop, the cyp82e5v2W422Stop, and cyp82e10 P381S Mutant Plants into the K346 Flue-CuredTobacco Cultivar

K346 SRC is a backcross-derived version of flue-cured tobacco cultivarK346 carrying introduced null mutations in three genes (CYP82E4 (SEQ IDNO: 5), CYP82E5 (SEQ ID NO: 8), and CYP82E10 (SEQ ID NO: 9)) previouslydocumented to encode for nicotine demethylase enzymes (Lewis et al.,2010). The introduced mutations in CYP82E4 (cyp82e4 W329Stop (SEQ ID NO:1)) and CYP82E5 (cyp82e5v2 W422Stop (SEQ ID NO: 2)) encode for prematurestop codons which render the genes non-functional. The introducedmutation in CYP82E10 (cyp82e10 P381S (SEQ ID NO: 11)) does not encodefor a premature stop codon, but does render the gene productnon-functional for converting nicotine to nornicotine (Lewis et al.,2010). When in homozygous condition, the three mutations result intobacco plants with (1) reduced genetic capacity to demethylate nicotineto form nornicotine, and (2) a corresponding diminished potential toaccumulate N-nitrosonornicotine (NNN), a potent carcinogen found in manytobacco products. The mutation in CYP82E4 (cyp82e4 W329Stop (SEQ ID NO:1)) also provides phenotypic stability for the “nicotine conversion”trait and eliminates the requirement to utilize the cumbersome “LC”method for reducing levels of nornicotine in tobacco cultivars.

The original tobacco cultivar K346 is a fertile inbred line. CMS K346 isa cytoplasmic male-sterile version of the original K346. To develop K346SRC, an individual plant of K346 is initially pollinated with a plant ofthe genetic background DH98-325-6 carrying mutations in each of thethree nicotine demethylase genes. F₁ individuals originating from thiscross and heterozygous for each mutation are backcrossed to therecurrent parent, K346, to produce BC₁F₁ progeny. BC₁F₁ progeny arescreened using genotyping methodologies to identify individualsheterozygous for all three mutations. A single triple heterozygous BC₁F₁plant is backcrossed to K346 to produce BC₂F₁ progeny. The process ofbackcrossing and identification of individuals heterozygous for allthree mutations is repeated through the BC₃F₁, BC₄F₁, BC₅F₁, BC₆F₁, andBC₇F₁ stages. At the BC₇F₁ stage, individuals heterozygous for all threemutations from each pedigree are self-pollinated to produce BC₇F₂ seed.A large number of BC₇F₂ progeny from each pedigree are genotyped toidentify individuals homozygous for all three mutations. A single BC₇F₂plant homozygous for all three mutations is self-pollinated to produce aBC₇F₃ family (K346 SRC) in which the wild-type CYP82E4, CYP82E5v2, andCYP82E10 alleles of K346 were replaced by the mutant (cyp82e4 W329Stop,cyp82e5v2 W422Stop, and cyp82e10 P381S alleles).

The male-sterile (CMS) version of K346 SRC (CMS K346 SRC) is produced bycrossing a plant of CMS K346 as a female with pollen of K346 SRC toproduce male-sterile plants heterozygous for all three mutations. Asingle male-sterile plant resulting from this cross is then subsequentlybackcrossed as a female to K346 SRC to produce progeny that aresegregating for individuals homozygous for all three mutations. Triplehomozygous individuals are identified by DNA genotyping to produce theCMS K346 SRC line. Because the line is male-sterile, it is maintainedvia pollination with K346 SRC.

Commercial K346 SRC is produced by pollinating plants of CMS K346 SRCwith pollen of K346 SRC.

Testing of K346 SRC

K346 SRC is evaluated for cured leaf chemistry, yield, and physicalquality at three North Carolina field research locations during 2012(Oxford, Rocky Mount, Kinston). K346 is included for comparison. Theexperimental design at each location is a randomized complete blockdesign with four replications. Experimental units are single 20-plantplots. Plots are harvested and flue-cured. Plot weights are used todetermine per acre yields. Cured leaf is evaluated by a former USDAtobacco grader. Fifty gram composite leaf samples are collected fromeach plot and analyzed for percent nicotine, nornicotine, anatabine,anabasine, and percent nicotine conversion using gas chromatographyequipment.

Comparisons using the least significant difference (LSD) test indicatedthat K346 SRC has significantly (P<0.05) lower levels of nornicotine andpercent nicotine conversion relative to K346 (Table 1). K346 and K346SRC are not significantly different from each other for percentnicotine, yield, or cured leaf quality indices.

Example 3 Breeding of Homozygous cyp82e4 W329Stop, the cyp82e5v2W422Stop, and cyp82e10 P381S Mutant Plants into the NC196 Flue-CuredTobacco Cultivar

Hybrid cultivar NC196 SRC is a backcross-derived version of flue-curedtobacco cultivar NC196 carrying introduced deleterious mutations inthree genes (CYP82E4 (SEQ ID NO: 5), CYP82E5 (SEQ ID NO: 8), andCYP82E10 (SEQ ID NO: 9)) previously documented to encode for nicotinedemethylase enzymes (Lewis et al., 2010). The introduced mutations inCYP82E4 (cyp82e4 W329Stop (SEQ ID NO: 1)) and CYP82E5 (cyp82e5v2W422Stop (SEQ ID NO: 2)) encode for premature stop codons which renderthe genes non-functional. The introduced mutation in CYP82E10 (cyp82e10P381S (SEQ ID NO: 11)) does not encode for a premature stop codon, butdoes render the gene product non-functional for converting nicotine tonornicotine (Lewis et al., 2010). When in homozygous condition, thethree mutations result in tobacco plants with (1) reduced geneticcapacity to demethylate nicotine to form nornicotine, and (2) acorresponding diminished potential to accumulate N-nitrosonornicotine(NNN), a potent carcinogen found in many tobacco products. The mutationin CYP82E4 (cyp82e4 W329Stop (SEQ ID NO: 1)) also provides phenotypicstability for the “nicotine conversion” trait and eliminates therequirement to utilize the cumbersome “LC” method for reducing levels ofnornicotine in tobacco cultivars.

The original tobacco cultivar NC196 is a hybrid generated by pollinatingplants of a male-sterile breeding line CMS NCTG-61 with pollen producedby fertile breeding line NC1562-1. To develop hybrid cultivar NC196 SRC,individual plants of fertile NCTG-61 and NC1562-1 are first pollinatedwith a plant of the genetic background DH98-325-6 carrying mutations ineach of the three nicotine demethylase genes. F₁ individuals originatingfrom each cross and heterozygous for each mutation are backcrossed tothe recurrent parents (NCTG-61 or NC1562-1) to produce BC₁F₁ progenies.

BC₁F₁ progenies are screened using genotyping methodologies to identifyindividuals heterozygous for all three mutations. A single BC₁F₁ plantfrom each pedigree is backcrossed to either NCTG-61 or NC1562-1,respectively, to produce BC₂F₁ progenies. The process of backcrossingand identification of individuals heterozygous for all three mutationsis repeated through the BC₃F₁, BC₄F₁, BC₅F₁, BC₆F₁, and BC₇F₁ stages. Atthe BC₇F₁ stage, individuals heterozygous for all three mutations fromeach pedigree are self-pollinated to produce BC₇F₂ seed. A large numberof BC₇F₂ progeny from each pedigree are genotyped to identifyindividuals homozygous for all three mutations. BC₇F₂ plants homozygousfor all three mutations are self-pollinated to produce BC₇F₃ families(NCTG-61 SRC and NC1562-1 SRC) in which the wild-type CYP82E4,CYP82E5v2, and CYP82E10 alleles of NCTG-61 and NC1562-1 are replaced bythe mutant (cyp82e4 W329Stop, cyp82e5v2 W422Stop, and cyp82e10 P381Salleles).

The female parental line of NC196, CMS NCTG-61, is cytoplasmic malesterile which causes pollen to not be produced. To develop CMS NCTG-61SRC, a plant of CMS NCTG-61 is crossed with NCTG-61 SRC to producemale-sterile plants heterozygous for all three mutations. A singlemale-sterile plant resulting from this cross is backcrossed as a femaleto NCTG-61 SRC to produce progeny that are segregating for individualshomozygous for all three mutations. Triple homozygous individuals areidentified by DNA genotyping to produce the CMS NCTG-61 SRC line.Because the line is male-sterile, it is maintained via pollination withNCTG-61 SRC.

Hybrid cultivar NC196 SRC is produced by pollinating plants of CMSNCTG-61 SRC with pollen of NC1562-1 SRC.

Testing of Hybrid Cultivar NC196 SRC

Hybrid cultivar NC196 SRC is evaluated for cured leaf chemistry, yield,and physical quality at three North Carolina field research locationsduring 2012 (Oxford, Rocky Mount, Kinston). Hybrid cultivar NC196 isincluded for comparison. The experimental design at each location is arandomized complete block design with four replications. Experimentalunits are single 20-plant plots. Plots are harvested and flue-cured.Plot weights are used to determine per acre yields. Cured leaf isevaluated by a former USDA tobacco grader. Fifty gram composite leafsamples are collected from each plot and analyzed for percent nicotine,nornicotine, anatabine, anabasine, and percent nicotine conversion usinggas chromatography equipment.

Comparisons using the least significant difference (LSD) test indicatesthat hybrid cultivar NC196 SRC has significantly (P<0.05) lower levelsof nornicotine and percent nicotine conversion relative to NC196 (Table1). NC196 and hybrid cultivar NC196 SRC are not significantly differentfrom each other for percent nicotine, yield, or cured leaf qualityindices,

TABLE 1 Means for Selected SRC Entries and EX12-122 entries averagedover three NC environments. Nicotine Nicotine Nornicotine AnabasineAnatabine Alkaloids Sugars Conversion Grade (% dry (% dry (% dry (% dry(% dry (% (% dry Genotype Yield Cwt Acre Index weight) weight) weight)weight) weight) reducing) weight) NC 2326 2269 134.73 3089.22 80.6 3.5810.064 0.027 0.203 3.876 13.59 1.76 NC 95 2519 139.44 3589.48 83.0 3.6600.068 0.033 0.284 4.046 12.22 1.83 NCTG-61 2776 141.71 3957.14 83.83.659 0.069 0.032 0.258 4.019 14.16 1.85 NCTG-61 2720 142.63 3895.7783.3 3.983 0.022 0.032 0.240 4.277 14.24 0.55 SRC K346 2757 141.913948.65 84.2 3.316 0.065 0.024 0.155 3.560 14.43 1.93 CMS K346 2590140.46 3656.09 83.2 3.425 0.019 0.027 0.187 3.659 10.85 0.56 SRC K3263076 143.01 4447.85 83.7 3.168 0.069 0.025 0.178 3.441 14.24 2.13 K326SRC 3048 143.99 4436.36 84.1 3.215 0.023 0.022 0.159 3.418 15.41 0.70NC1562-1 3052 144.81 4463.77 84.2 2.886 0.052 0.022 0.159 3.120 15.881.79 NC1562-1 3091 142.94 4454.60 83.8 2.950 0.016 0.022 0.170 3.15815.73 0.55 SRC NC 196 3042 142.76 4392.61 83.4 3.215 0.070 0.030 0.2533.568 14.24 2.14 NC196 SRC 3155 143.23 4551.78 83.9 3.508 0.020 0.0270.210 3.765 15.56 0.57 LSD 0.05 246 5.47 401.23 2.7 0.343 0.010 0.0030.027 0.367 2.22 0.37

Deposit Information

A deposit of the proprietary inbred plant lines and hybrid disclosedabove and recited in the appended claims have been made with AmericanType Culture Collection (ATCC), 10801 University Boulevard, Manassas,Va. 20110. The date of deposit for K326 SRC, CMS K326 SRC, K346 SRC, CMSK346 SRC, NC1562-1 SRC, NC196 SRC, and CMS NCTG-61 SRC was Jan. 17,2013. The date of deposit for NCTG-61 SRC was May 7, 2013. The depositsof 2500 seeds for each variety was taken from the same depositsmaintained since prior to the filing date of this application. Uponissuance of a patent, all restrictions upon the deposits will beremoved, and the deposits are intended to meet all of the requirementsof 37 C.F.R. §1.801-1.809. The ATCC has issued the following accessionnumbers: ATCC Accession No. PTA-13455 for K326 SRC; ATCC Accession No.PTA-13454 for CMS K326 SRC; ATCC Accession No. PTA-13453 for K346 SRC;ATCC Accession No. PTA-13452 for CMS K346 SRC; ATCC Accession No.PTA-13457 for NC1562-1 SRC; ATCC Accession No. PTA-13456 for CMS NCTG-61SRC; ATCC Accession No. PTA-120314 for NCTG-61 SRC; and ATCC AccessionNo. PTA-13458 for hybrid NC196 SRC. These deposits will be maintained inthe depository for a period of 30 years, or 5 years after the lastrequest, or for the effective life of the patent, whichever is longer,and will be replaced as necessary during that period.

What is claimed is:
 1. Cured tobacco material or a tobacco productprepared therefrom, wherein said cured tobacco material is from atobacco plant, or part thereof, of a hybrid, a cultivar, or a progenyhybrid produced from said cultivar, said hybrid is NC196 SRC, saidcultivar is selected from the group consisting of K326 SRC, CMS K326SRC, K346 SRC, CMS K346 SRC, NC1562-1 SRC, NCTG-61 SRC, and CMS NCTG-61SRC; a representative sample seed of said K326 SRC, CMS K326 SRC, K346SRC, CMS K346 SRC, NC1562-1 SRC, NC196 SRC, NCTG-61 SRC, and CMS NCTG-61SRC having been deposited with the ATCC under ATCC Accession Nos.PTA-13455, PTA-13454, PTA-13453, PTA-13452, PTA-13457, PTA-13458,PTA-120314, and PTA-13456, respectively.
 2. The tobacco product of claim1, wherein said tobacco product is selected from the group consisting ofa pipe tobacco, a cigar tobacco, a cigarette tobacco, a chewing tobacco,a leaf tobacco, a shredded tobacco, and a cut tobacco.
 3. The tobaccoproduct of claim 1, wherein said tobacco product is selected from thegroup consisting of a cigarillo, a non-ventilated recess filtercigarette, a vented recess filter cigarette, a cigar, snuff, and achewing tobacco.
 4. The tobacco product of claim 3, wherein said tobaccoproduct has an amount of nornicotine of less than about 3 mg/g.
 5. Thetobacco product of claim 4, wherein said amount of nicotine is selectedfrom the group consisting of 3.0 mg/g, 2.5 mg/g, 2.0 mg/g, 1.5 mg/g, 1.0mg/g, 750 μg/g, 500 pg/g, 250 pg/g, 100 pg/g, 75 pg/g, 50 pg/g, 25 pg/g,10 pg/g, 7.0 pg/g, 5.0 pg/g, 4.0 pg/g, 2.0 pg/g, 1.0 pg/g, 0.5 pg/g, 0.4pg/g, 0.2 pg/g, 0.1 pg/g, 0.05 pg/g, 0.01 pg/g, and undetectable.
 6. Thetobacco product of claim 3, wherein said tobacco product has an amountof N′-nitrosonornicotine (NNN) of less than about 10 pg/g.
 7. Thetobacco product of claim 6, wherein said amount of NNN is selected fromthe group consisting of 10 pg/g, 7.0 pg/g, 5.0 pg/g, 4.0 pg/g, 2.0 pg/g,1.0 pg/g, 0.5 pg/g, 0.4 pg/g, 0.2 pg/g, 0.1 pg/g, 0.05 pg/g, 0.01 pg/g,and undetectable.
 8. The tobacco product of claim 1, wherein said curedtobacco material is at least about 5% by dry weight of the total tobaccomaterial in said tobacco product.
 9. The tobacco product of claim 8,wherein said cured tobacco material has a dry weight percentage of saidtotal tobacco material selected from the group consisting of at leastabout 10%, at least about 20%, at least about 30%, at least about 40%,at least about 50%, at least about 60%, at least about 70%, at leastabout 80%, at least about 90%, and at least about 95%.
 10. A method ofproducing a tobacco product, comprising: providing cured tobaccomaterial from a tobacco plant, or part thereof, of a hybrid, a cultivar,or a hybrid produced from said cultivar, wherein said hybrid is NC196SRC, said cultivar is selected from the group consisting of CMS K326SRC, CMS K326 SRC, K346 SRC, CMS K346 SRC, NC1562-1 SRC, NCTG-61 SRC,and CMS NCTG-61 preparing a tobacco product from said cured tobaccomaterial, a representative sample seed of said K326 SRC, CMS K326 SRC,K346 SRC, CMS K346 SRC, NC1562-1 SRC, NC196 SRC, NCTG-61 SRC, and CMSNCTG-61 SRC cultivars having been deposited with the ATCC under ATCCAccession Nos. PTA-13455, PTA-13454, PTA-13453, PTA-13452, PTA-13457,PTA-13458, PTA-120314, and PTA-13456, respectively.
 11. A seed oftobacco cultivar selected from the group consisting of K326 SRC, CMSK326 SRC, K346 SRC, CMS K346 SRC, NC1562-1 SRC, NC196 SRC, and CMSNCTG-61 SRC; a representative sample seed of said K326 SRC, CMS K326SRC, K346 SRC, CMS K346 SRC, NC1562-1 SRC, NC196 SRC, NCTG-61 SRC, andCMS NCTG-61 SRC tobacco cultivars having been deposited with the ATCCunder ATCC Accession Nos. PTA-13455, PTA-13454, PTA-13453, PTA-13452,PTA-13457, PTA-13458, PTA-120314, and PTA-13456, respectively.
 12. Atobacco plant, or part thereof, produced from said seed of claim
 11. 13.The part of the plant of claim 12, wherein said part is selected fromthe group consisting of a harvested leaf, a pollen, an ovule, an embryo,a cotyledon, a hypocotyl, a meristematic cell, a protoplast, a root, aroot tip, a pistil, an anther, a flower, a shoot, a stem, a pod, apetiole, and combinations thereof.
 14. A harvested leaf of said tobaccoplant of claim
 12. 15. The harvested leaf of claim 14, wherein said leafhas a reduced amount of nornicotine and/or N′-nitrosonomicotine (NNN).16. The harvested leaf of claim 15, wherein said reduced amount ofnornicotine and/or N-nitrosonornicotine (NNN) is reduced in a smokestream produced from said leaf.
 17. An F₁ progeny plant of a tobaccocultivar selected from the group consisting of K326 SRC, CMS K326 SRC,K346 SRC, CMS K346 SRC, NC1562-1 SRC, NCTG-61 SRC, and CMS NCTG-61 SRC;a representative sample seed of said K326 SRC, CMS K326 SRC, K346 SRC,CMS K346 SRC, NC1562-1 SRC, NCTG-61 SRC, and CMS NCTG-61 SRC tobaccocultivars having been deposited with the ATCC under ATCC Accession Nos.PTA-13455, PTA-13454, PTA-13453, PTA-13452, PTA-13457, PTA-120314, andPTA-13456, respectively.
 18. The F₁ progeny plant of claim 17, whereinsaid F₁ plant is male sterile.